circCPA4 induces malignant behaviors of prostate cancer via miR-491-5p/ SHOC2 feedback loop

ABSTRACT Objective: circCPA4 has been defined to be an oncogenic gene. This study examined whether circCPA4 regulates Prostate Cancer (PC) development and revealed its molecular mechanism. Methods: PC tissues and PC cell lines were collected, in which circCPA4/miR-491-5p/SHOC2 levels were evaluated by RT-qPCR and immunoblot. Colony formation assay and EdU assay assessed cell proliferation, flow cytometry measured apoptosis, and Transwell assessed invasion and migration. Ki-67, cleaved caspase-3, E-cadherin, and N-cadherin were evaluated by immunoblot. Based on the luciferase reporter assay and RIP assay the authors investigated the targeting relationship between circCPA4/miR-491-5p/SHOC2. The effect of circCPA4 on tumor growth was evaluated by xenotransplantation in nude mice. Results: circCPA4 and SHOC2 levels were abundant while miR-491-5p expression was low in PC. Loss of circCPA4 decreased the proliferation and EdU-positive rate of PC cells, enhanced apoptosis, and inhibited invasion, migration, and EMT. Upregulation of circCPA4 forced the malignant behaviors of PC cells, and this promotion could be abolished when miR-491-5p was overexpressed or SHOC2 was silenced. CircCAP4 competitively decoyed miR-491-5p mediating SHOC2 expression. circCAP4 suppression inhibited PC tumor growth. Conclusion: circCAP4 acts as a novel oncogenic factor in PC, accelerating the malignant behavior of PC cells via miR-491-5p/SHOC2 interaction. This novel ceRNA axis may be a potential target for PC drug development and targeted therapy in the future.

The association of polypoid growth pattern with lymphnode involvement in endometrioid type endometrial adenocarcinoma

Background: Tumor size is an independent predictor of lymph node metastasis and survival in the endometrioid type endometrial adenocarcinoma (EC). However, some of the ECs tend to grow towards the cavity in the polypoid pattern, which can reach very large sizes. In this study, we aimed to analyze the association of growing in the polypoid pattern of the tumor with the proportion of lymph node metastasis and extrauterine tumor spread. Methods: Four hundred seven patients were analyzed retrospectively. The effect of tumor size, tumor growing pattern, myometrial invasion, grade, and lymphovascular space invasion on the lymph node metastasis and extrauterine tumor spread were investigated. Statistical analysis consisted of unpaired t?tests for parametric data and Mann Whitney?U test for non?parametric data, whereas the Chi?square test for categorical variables. Logistic Regression, Cox Regression and multivariate analysis were used to estimate the risk predictors. Results: No association was found between the growing in polypoid pattern and lymph node metastasis (P > 0.05). In the analysis of endometrioid type EC patients who had myometrial invasion less than � as a subgroup, no association was found between the growing pattern and lymph node metastasis and extrauterine disease. Tumor size was found to be a statistically significant predictor of lymph node metastasis and extrauterine disease (P < 0.05). Conclusions: Lymphovascular space invasion, grade, and myometrial invasion are associated with a higher proportion of lymph node metastasis. The polypoid growth pattern of the tumor does not correlate with any histopathological parameters

MicroRNA-150-5p inhibits the proliferation and invasion of human larynx epidermiod cancer cells though regulating peptidyl-prolyl cis/trans isomerase

Abstract Objective This study aimed to investigate the molecular mechanism of miR-150-5p regulating the malignant biological behavior of Human Epidermoid cancer cell (HEp-2) by targeting peptidyl-prolyl cis/trans isomerase NIMA-Interacting-1 (PIN1). Methods Firstly, qRT-PCR and Western blot were adopted to detect the expression levels of miR-150-5p and PIN1 in cancer tissue and paracancerous tissues of patients with LSCC, and those in human bronchial epithelial cells (16 HBE) and HEp-2. Next, the targeted relationship between miR-150-5p and PIN1 was assessed by bioinformatics website and dual-luciferase reporter assay, followed by their correlation analysis. Besides, after interfering with miR-150-5p or PIN1 expression in HEp-2 cells, CCK-8, cell colony formation assay, and transwell assay were utilized to detect the proliferation, viability, and invasion of cells, respectively. Subsequently, the protein levels of MMP-2, MMP-9, and EMT-related proteins in HEp-2 cells were checked by Western blot. Results Expression of miR-150-5p was down-regulated in LSCC tissues and HEp-2 cells. Moreover, miR-150-5p suppressed proliferation and invasion of HEp-2 cells, affected protein expression related to MMP and EMT, thereby inhibiting development of cancer. The expression of PIN1 was significantly increased in cancer tissues and HEp-2 cells, and there was a targeted relationship and negative correlation between miR-150-5p and PIN1 in cancer tissue. However, overexpression of PIN1 could reverse the effect of miR-150-5p on the proliferation and invasion of HEp-2 cells. Conclusion In a nutshell, there is a targeted relationship between PIN1 and miR-150-5p. Besides, miR-150-5p can inhibit the proliferation and invasion of HEp-2 cells by regulating the expression of PIN1. Level of evidence 3.

Doublecortin-like kinase 1 activates Hippo pathway to promote migration, invasion and proliferation of pancreatic cancer cells / 中华肿瘤杂志

Objective: To explore the mechanism of Doublecortin-like kinase 1 (DCLK1) in promoting cell migration, invasion and proliferation in pancreatic cancer. Methods: The correlation between DCLK1 and Hippo pathway was analyzed using TCGA and GTEx databases and confirmed by fluorescence staining of pancreatic cancer tissue microarrays. At the cellular level, immunofluorescence staining of cell crawls and western blot assays were performed to clarify whether DCLK1 regulates yes associated protein1 (YAP1), a downstream effector of the Hippo pathway. Reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR) was used to analyze the expressions of YAP1 binding transcription factor TEA-DNA binding proteins (TEAD) and downstream malignant behavior-promoting molecules CYR61, EDN1, AREG, and CTGF. Transwell test of the DCLK1-overexpressing cells treated with the Hippo pathway inhibitor Verteporfin was used to examine whether the malignant behavior-promoting ability was blocked. Analysis of changes in the proliferation index of experimental cells used real-time label-free cells. Results: TCGA combined with GTEx data analysis showed that the expressions of DCLK1 and YAP1 molecules in pancreatic cancer tissues were significantly higher than those in adjacent tissues (P<0.05). Moreover, DCLK1was positively correlated with the expressions of many effectors in the Hippo pathway, including LATS1 (r=0.53, P<0.001), LATS2 (r=0.34, P<0.001), MOB1B (r=0.40, P<0.001). In addition, the tissue microarray of pancreatic cancer patients was stained with multicolor fluorescence, indicated that the high expression of DCLK1 in pancreatic cancer patients was accompanied by the up-regulated expression of YAP1. The expression of DCLK1 in pancreatic cancer cell lines was analyzed by the CCLE database. The results showed that the expression of DCLK1 in AsPC-1 and PANC-1 cells was low. Thus, we overexpressed DCLK1 in AsPC-1 and PANC-1 cell lines and found that DCLK1 overexpression in pancreatic cancer cell lines promoted YAP1 expression and accessible to the nucleus. In addition, DCLK1 up-regulated the expression of YAP1 binding transcription factor TEAD and increased the mRNA expression levels of downstream malignant behavior-promoting molecules. Finally, Verteporfin, an inhibitor of the Hippo pathway, could antagonize the cell's malignant behavior-promoting ability mediated by high expression of DCLK1. We found that the number of migrated cells with DCLK1 overexpressing AsPC-1 group was 68.33±7.09, which was significantly higher than 22.00±4.58 of DCLK1 overexpressing cells treated with Verteporfin (P<0.05). Similarly, the migration number of PANC-1 cells overexpressing DCLK1 was 65.66±8.73, which was significantly higher than 37.00±6.00 of the control group and 32.33±9.61 of Hippo pathway inhibitor-treated group (P<0.05). Meanwhile, the number of invasive cells in the DCLK1-overexpressed group was significantly higher than that in the DCLK1 wild-type group cells, while the Verteporfin-treated DCLK1-overexpressed cells showed a significant decrease. In addition, we monitored the cell proliferation index using the real-time cellular analysis (RTCA) assay, and the proliferation index of DCLK1-overexpressed AsPC-1 cells was 0.66±0.04, which was significantly higher than 0.38±0.01 of DCLK1 wild-type AsPC-1 cells (P<0.05) as well as 0.05±0.03 of DCLK1-overexpressed AsPC1 cells treated with Verteporfin (P<0.05). PANC-1 cells showed the same pattern, with a proliferation index of 0.77±0.04 for DCLK1-overexpressed PANC-1 cells, significantly higher than DCLK1-overexpressed PANC1 cells after Verteporfin treatment (0.14±0.05, P<0.05). Conclusion: The expression of DCLK1 is remarkably associated with the Hippo pathway, it promotes the migration, invasion, and proliferation of pancreatic cancer cells by activating the Hippo pathway.

LncRNA DRAIC regulates the proliferation, apoptosis, migration and invasion of lung adenocarcinoma cells by targeting let-7i-5p / 中华肿瘤杂志

Objective: To investigate the effects of lncRNA DRAIC on proliferation, apoptosis, migration and invasion of lung adenocarcinoma cells and its mechanism. Methods: Reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR) was used to detect the expression of DRAIC in lung cancer tissues and corresponding adjacent normal tissues of 40 patients with lung adenocarcinoma who underwent surgery in Tangshan People's Hospital from 2019 to 2020. Lung adenocarcinoma cells A549 and H1299 were cultured in vitro and divided into si-NC group, si-DRAIC group, miR-NC group, let-7i-5p mimics group, si-DRAIC+ inhibitor-NC group, and si-DRAIC+ let-7i-5p inhibitor group. CCK-8 method and clone formation experiment were used to detect cell proliferation. Flow cytometry was used to detect cell apoptosis. Transwell array was used to detect the cell migration and invasion. Western blot was used to detect the protein expressions of Caspase-3, Caspase-9, Bcl-2 and Bax. The double luciferase reporter gene experiment was used to verify the regulatory relationship between DRAIC and let-7i-5p. Independent sample t test was used for comparison between two groups, one-way ANOVA was used for comparison between multiple groups, and Pearson correlation analysis was used for correlation analysis. Results: Compared with adjacent tissues, the expression level of DRAIC in lung adenocarcinoma tissues increased (P<0.05), but the expression level of let-7i-5p decreased (P<0.05). The expression levels of DRAIC and let-7i-5p in lung adenocarcinoma tissues were negatively correlated (r=-0.737, P<0.05). The absorbance value of A549 and H1299 cells in the si-DRAIC group at 48, 72 and 96 hours were lower than those in the si-NC group (P<0.05), the number of clones formed [(91.00±6.08 vs. 136.67±6.51); (50.67±1.53 vs. 76.67±4.51)], the number of migration [(606.67±31.34 vs. 960.00±33.06); (483.33±45.96 vs. 741.67±29.67)], the number of invasion [(185.00±8.19 vs. 447.33±22.05); (365.00±33.87 vs. 688.00±32.97)] were lower than those in the si-NC group (P<0.05). However, the apoptosis rates of cells [(13.43±2.79)% vs. (4.53±0.42)%; (23.77±1.04)% vs. (6.60±1.42)%] were higher than those in the si-NC group (P<0.05). The protein expressions of Caspase-3, Caspase-9 and Bax in si-DRAIC group were higher than those in si-NC group, and the protein expression of Bcl-2 was lower than that in si-NC group (P<0.05). DRAIC is located in the cytoplasm. DRAIC targeted and negatively regulated the expression of let-7i-5p. The absorbance values of A549 and H1299 cells in the let-7i-5p mimics group at 48, 72 and 96 hours were lower than those in the miR-NC group (P<0.05), the number of clones formed [(131.33±14.47 vs. 171.33±6.11); (59.33±4.93 vs. 80.33±7.09)], the number of migration [(137.67±3.06 vs. 579.33±82.03); (425.00±11.14 vs. 669.33±21.13)], the number of invasion [(54.00±4.36 vs. 112.67±11.59); (80.00±4.58 vs. 333.33±16.80)] were lower than those in the miR-NC group (P<0.05). However, the apoptosis rates of cells [(14.57±1.10)% vs. (6.97±1.11)%; (23.97±0.42)% vs. (7.07±1.21)%] were higher than those in the miR-NC group (P<0.05). The protein expressions of Caspase-3, Caspase-9 and Bax in let-7i-5p mimics group were higher than those in miR-NC group, and the protein expression of Bcl-2 was lower than that in miR-NC group (P<0.05). The absorbance values of A549 and H1299 cells in the si-DRAIC+ let-7i-5p inhibitor group at 48, 72 and 96 hours were higher than those in the si-DRAIC+ inhibitor-NC group (P<0.05), the number of clones formed [(82.00±5.29 vs. 59.00±5.57); (77.67±4.93 vs. 41.33±7.57)], the number of migration [(774.33±35.81 vs. 455.67±19.04); (569.67±18.72 vs. 433.67±16.77)], the number of invasion [(670.33±17.21 vs. 451.00±17.52); (263.67±3.06 vs. 182.33±11.93)] were higher than those in the si-DRAIC+ inhibitor-NC group (P<0.05). However, the apoptosis rates of cells [(7.73±0.45)% vs. (19.13±1.50)%; (8.00±0.53)% vs. (28.40±0.53)%] were lower than those in the si-NC group (P<0.05). The protein expressions of Caspase-3, Caspase-9 and Bax in si-DRAIC+ let-7i-5p inhibitor group were higher than those in si-DRAIC+ inhibitor-NC group, and the protein expression of Bcl-2 was lower than that in si-DRAIC+ inhibitor-NC group (P<0.05). Conclusion: DRAIC is highly expressed in lung adenocarcinoma, and DRAIC promotes the proliferation, migration and invasion of lung adenocarcinoma cells and inhibits apoptosis by targeting let-7i-5p.

Research Progress of Pseudogenes in Hepatocellular Carcinoma / 肿瘤防治研究

Pseudogenes were initially thought to have no function and were called by aliases, such as "junk genes." With the emergence of large-scale genomics projects and more and more experimental studies, pseudogenes have been shown to play an important role in the occurrence and development of solid tumors, especially playing an important regulatory role in the occurrence and develepment of liver cancer, such as regulating the proliferation, apoptosis, invasion, metastasis, and immunity of liver cancer cells. Recent studies showed that pseudogenes can act as regulators of oncogenes and tumor suppressors in hepatocellular carcinoma (HCC) and can thus serve as prognostic markers and even therapeutic targets for this cancer type. In this review, we systematically summarize the mechanisms and functions of different pseudogenes in HCC and present their future prospects as therapeutic targets.

Development and validation of a nomogram model for preoperative prediction of hepatocellular carcinoma with microvascular invasion / 中华肝胆外科杂志

Objective:To develop and validate a nomogram model for predicting microvascular invasion (MVI) in hepatocellular carcinoma (HCC) based on preoperative enhanced computed tomography imaging features and clinical data.Methods:The clinical data of 210 patients with HCC undergoing surgery in the Second Affiliated Hospital of Anhui Medical University from May 2018 to May 2022 were retrospectively analyzed, including 172 males and 38 females, aged (59±10) years old. Patients were randomly divided into the training group ( n=147) and validation group ( n=63) by systematic sampling at a ratio of 7∶3. Preoperative enhanced computed tomography imaging features and clinical data of the patients were collected. Logistic regression was conducted to analyze the risk factors for HCC with MVI, and a nomogram model containing the risk factors was established and validated. The diagnostic efficacy of predicting MVI status in patients with HCC was assessed by receiver operating characteristic (ROC) curve, calibration curves, decision curve analysis (DCA), and clinical impact curve (CIC) of the subjects in the training and validation groups. Results:The results of multifactorial analysis showed that alpha fetoprotein ≥400 μg/ml, intra-tumor necrosis, tumor length diameter ≥3 cm, unclear tumor border, and subfoci around the tumor were independent risk factors predicting MVI in HCC. A nomogram model was established based on the above factors, in which the area under the curve (AUC) of ROC were 0.866 (95% CI: 0.807-0.924) and 0.834 (95% CI: 0.729-0.939) in the training and validation groups, respectively. The DCA results showed that the predictive model thresholds when the net return is >0 ranging from 7% to 93% and 12% to 87% in the training and validation groups, respectively. The CIC results showed that the group of patients with predictive MVI by the nomogram model are highly matched with the group of patients with confirmed MVI. Conclusion:The nomogram model based on the imaging features and clinical data could predict the MVI in HCC patients prior to surgery.

Effects of methyl methanesulfonate and UV sensitive gene clone 81 on hepatocellular carcinoma cells / 中华肝胆外科杂志

Objective:To detect the expression of methyl methanesulfonate and UV sensitive gene clone 81 (Mus81) in hepatocellular carcinoma(HCC) and to observe the effects of Mus81 on the migration, invasion and metastatic ability of human HCC cells.Methods:Thirty-two tissue specimens were selected from HCC tissues and corresponding paraneoplastic tissues of patients with HCC who underwent surgical resection in Guangzhou Red Cross Hospital Affiliated to Jinan University from January 2020 to June 2021. The expression levels of Mus81 in 32 HCC specimens, 374 HCC samples from the cancer genome atlas database, human normal liver cell line HL-7702 and human HCC cell lines JHH-7, Huh-7 and Hep3B were analyzed. Mus81 knockdown in JHH-7, Huh-7 and overexpressed in Hep3B HCC cell lines were constructed, and the effects of Mus81 on HCC cells were observed by scratch assay, Transwell migration and invasion assay and tail vein injection transfer assay in nude mice.Results:The expression of Mus81 was higher in HCC tissues or cell lines than which in paraneoplastic tissues or normal hepatocyte lines (all P<0.05). The migration rate, metastatic and invasive cell numbers of Mus81-knockdown Huh-7 HCC cells were 22.24%±2.16%, 49.04±5.62, 3.81±1.08, the negative control group were 26.89%±1.15%, 86.81±4.79, 19.78±3.30, and the differences between the two groups were statistically significant ( t=4.24, 26.59, 23.92, all P<0.01). The migration rate, metastatic and invasive cell numbers of Mus81-overexpressed Hep3B HCC cells were 80.57%±5.12%, 18.74±8.07, 33.81±8.44, which were significantly higher than those of the empty vector group 64.17%±7.20%, 10.96±5.32, 3.04±1.13, and the differences were statistically significant ( t=4.15, 4.18, 18.78, all P<0.01). Tail vein transfer experiments in nude mice showed that the total fluorescence expression, weight of metastatic tumors, and the metastatic rates in kidney, vertebral column, neck, axilla and subcutis in nude mice injected with Mus81-knockdown JHH-7 cells were significantly lower than those in the control group (all P<0.05). Conclusion:Mus81 gene expression is upregulated in HCC and promotes the migration, invasion and metastatic ability of HCC cells, suggesting that Mus81 may be a potential therapeutic target for HCC.

Performance of laminin γ2 in diagnosing hepatocellular carcinoma and in microvascular invasion / 中华肝胆外科杂志

Objective:To analyze the value of laminin γ2 (LAMC2) in the diagnosis of hepatocellular carcinoma (HCC) and the difference in patients with different types of microvascular invasion (MVI).Methods:A cohort of 100 patients with HCC who underwent surgical treatment at the Faculty of Hepato-Pancreato-Biliary Surgery, Chinese PLA General Hospital from January 2021 to March 2022 were prospectively enrolled. There were 80 males and 20 females, aged (55.7±11.1) years. The data of 17 patients with hepatic hemangioma without cirrhosis who underwent operation at the same hospital during the study period were collected to serve as the control group (6 males, 11 females), aged (42.8±9.8) years. LAMC2 in serum was determined by enzyme linked immunosorbent assay. The levels of alpha-fetoprotein (AFP) and LAMC2 were compared between the two groups, and receiver operating characteristic (ROC) curves were drawn to compare these two markers in the diagnosis of HCC. The LAMC2 of different MVI patients were compared.Results:The levels of LAMC2 and AFP were 1 334.2(838.9, 2 656.0) pg/ml and 19.0(4.6, 778.6) μg/L in the HCC group, which were significantly higher than 375.2(221.2, 691.7)pg/ml and 3.3(2.5, 3.5) μg/L in the control group ( Z=-4.32, -4.63, both P<0.001). The areas under the ROC curve were 0.829(95% CI: 0.748-0.892) for LAMC2 and 0.852(95% CI: 0.769-0.910) for AFP, and was 0.949(95% CI: 0.911-0.988) for using both in the diagnoses. The diagnostic efficacy of combining LAMC2 and AFP was significantly better than that of LAMC2 alone and AFP alone (area under ROC: Z=3.15, 3.07, P=0.002, 0.002). When the patients were divided into the M0 group (61 patients), the M1 Group (25 patients) and the M2 Group (14 patients) based on MVIs, the concentrations of LAMC2 were 1 168.6(834.3, 2 521.4) pg/ml, 942.2(614.0, 2 056.6) pg/ml and 3 128.4(1 852.7, 7 191.3) pg/ml, respectively. The level of LAMC2 in the M2 group was significantly higher than that in the M0 and M1 groups ( Z=-3.46, -3.32, P=0.001, 0.004). Conclusion:The diagnostic efficacy of LAMC2 combined with AFP for HCC was significantly higher than that of either LAMC2 alone or AFP alone. Serum LAMC2 levels were significant different among the groups of HCC patients with different types of MVI.

Preoperative prediction of vessel invasion in locally advanced gastric cancer based on venous phase enhanced CT radiomics and machine learning / 中华放射学杂志

Objective:To evaluate the value of preoperative prediction of vessel invasion (VI) of locally advanced gastric cancer by machine learning model based on the venous phase enhanced CT radiomics features.Methods:A retrospective analysis of 296 patients with locally advanced gastric cancer confirmed by pathology in the First Affiliated Hospital of Zhengzhou University from July 2011 to December 2020 was performed. The patients were divided into VI positive group ( n=213) and VI negative group ( n=83) based on pathological results. The data were divided into training set ( n=207) and test set ( n=89) according to the ratio of 7∶3 with stratification sampling. The clinical characteristics of patients were recorded, and the independent risk factors of gastric cancer VI were screened by multivariate logistic regression. Pyradiomics software was used to extract radiomic features from the venous phase enhanced CT images, and the minimum absolute shrinkage and selection algorithm (LASSO) was used to screen the features, obtain the optimal feature subset, and establish the radiomics signature. Four machine learning algorithms, including extreme gradient boosting (XGBoost), logistic, naive Bayes (GNB), and support vector machine (SVM) models, were used to build prediction models for the radiomics signature and the screened clinical independent risk factors. The efficacy of the model in predicting gastric cancer VI was evaluated by the receiver operating characteristic curve. Results:The degree of differentiation (OR=13.651, 95%CI 7.265-25.650, P=0.003), Lauren′s classification (OR=1.349, 95%CI 1.011-1.799, P=0.042) and CA199 (OR=1.796, 95%CI 1.406-2.186, P=0.044) were independent risk factors for predicting the VI of locally advanced gastric cancer. Based on the venous phase enhanced CT images, 864 quantitative features were extracted, and 18 best constructed radiomics signature were selected by LASSO. In the training set, the area under the curve (AUC) of XGBoost, logistic, GNB and SVM models for predicting gastric cancer VI were 0.914 (95%CI 0.875-0.953), 0.897 (95%CI 0.853-0.940), 0.880 (95%CI 0.832-0.928) and 0.814 (95%CI 0.755-0.873), respectively, and in the test set were 0.870 (95%CI 0.769-0.971), 0.877 (95%CI 0.788-0.964), 0.859 (95%CI 0.755-0.961) and 0.773 (95%CI 0.647-0.898). The logistic model had the largest AUC in the test set. Conclusions:The machine learning model based on the venous phase enhanced CT radiomics features has high efficacy in predicting the VI of locally advanced gastric cancer before the operation, and the logistic model demonstrates the best diagnostic efficacy.

Value of spectral CT based iodine concentration for the preoperative prediction of vascular invasion in gastric cancer / 中华放射学杂志

Objective:To investigate the value of spectral CT based iodine concentration (IC) parameters for preoperative prediction of lymphovascular invasion (LVI) in gastric cancer.Methods:Between January 2021 and November 2021, 266 patients diagnosed as gastric adenocarcinomas by endoscopy and undergoing gastrectomy at the Affiliated Cancer Hospital of Zhengzhou University were recruited prospectively. They were divided into LVI and non-LVI groups according to pathological reports. Triple phase contrasted enhanced CT scans, including arterial phase (AP), venous phase (VP) and delayed phase (DP) were performed on a spectral CT platform within one week before surgery. The IC of gastric cancer lesions at three enhanced phases were measured based on iodine maps, and the normalized IC (nIC) was calculated. The thickness of the tumor was measured. Clinicopathological features were collected, including ulceration, pathological tumor staging (pT), pathological node staging (pN), histodifferentiation, Lauren subtype, perineural invasion (PNI), positive node numbers and positive node ratio. Student′s t tes t or Mann-Whitney U test were used to compare the differences of continuous variables between the two groups, while Chi-square test or Fisher′s exact test was used for categorical data. Multivariable logistic regression analysis was used to screen independent risk factors of LVI, and to build a combined parameter based on risk factors. The receiver operating characteristic curve analysis was performed to determine the predictive efficacy of IC parameters and the combined parameter for LVI. DeLong′s test was used to compare the differences among different area under the curve (AUC). Results:There were statistical differences in tumor thickness, ulceration, pT, pN, histodifferentiation, positive node numbers, positive node ratio, Lauren subtype and PNI between LVI and non-LVI groups ( P<0.05). The values of IC VP, IC DP, nIC VP, nIC DP in LVI group were statistically higher than those in non-LVI group ( t=3.77, 4.23, 4.25, 6.12, all P<0.001), with the AUC (95%CI) of 0.674 (0.610-0.738), 0.677 (0.614-0.741), 0.731 (0.671-0.792), 0.700 (0.636-0.764) for predicting LVI, respectively. Multivariable logistic regression analysis revealed that tumor thickness (OR=1.148, 95%CI 1.085-1.237, P<0.001) and nIC VP (OR=209.904, 95%CI 14.874-644.362, P<0.001) were independent predictors for LVI, the combined parameter incorporating these two factors yielded an AUC (95%CI) of 0.790 (0.736-0.937), which was statistically higher than any single parameter of IC VP, IC DP, nIC VP and nIC DP ( Z=3.07, 3.29, 2.10, 2.60, P=0.002, 0.001, 0.036, 0.009). Conclusion:The IC and nIC values of gastric cancer lesions derived from the VP and DP on spectral CT can effectively predict LVI status in gastric adenocarcinomas, and the combination of nIC VP and tumor thickness can further improve the predictive efficacy.

Preoperative breast MRI combined with axillary ultrasound for the prediction of lymphovascular invasion in invasive ductal carcinoma of the breast / 中华放射学杂志

Objective:To investigate the value of preoperative breast MRI combined with axillary ultrasound in predicting lymphovascular invasion (LVI) of breast invasive ductal carcinoma.Methods:The clinical, pathological and imaging features of 160 female patients [age 25-74(49±10)years] with breast invasive ductal carcinoma from March 2014 to December 2017 in Shanxi Cancer Hospital were retrospectively analyzed. According to the LVI status determined by postoperative pathology, 160 patients were divided into LVI positive group (56 cases) and LVI negative group (104 cases). The clinical characteristics, pathological characteristics and imaging features of LVI positive group and LVI negative group were compared by the independent t test, Mann-Whitney U test or χ 2 test. Multivariate logistic regression analysis was performed to identify independent predictors for predicting LVI and construct a predictive model. The receiver operating characteristic (ROC) curve and area under the curve (AUC) was used to evaluate the discrimination of the prediction model, and the Hosmer-Lemeshow test was used to evaluate its calibration. Results:There was no significant difference in age, menopausal status, estrogen receptor, progesterone receptor, human epidermal growth factor 2, Ki67 index and molecular subtype between LVI positive group and negative group ( P>0.05). Tumor size, peritumoral edema, adjacent vessel sign, multifocality or multicentricity, peritumoral maximum-apparent diffusion coefficient (ADC), peritumour-tumour ADC ratio, MRI axillary lymph node status and ultrasound axillary lymph node status between LVI positive group and LVI negative group showed significantly statistical difference ( P<0.05). Variables with significant difference in the univariate analysis were entered into multivariate logistic regression analysis to explore predictors for LVI. Peritumoral edema (OR=3.367, 95%CI 1.382-8.201, P=0.008), multifocality or multicentricity (OR=4.026, 95%CI 1.268-12.776, P=0.018), high peritumoral-tumor ADC ratio (OR=7.321, 95%CI 2.226-24.079, P=0.001) and positive ultrasound axillary lymph node (OR=6.779, 95%CI 2.819-16.303, P<0.001) were independent predictors for predicting LVI. A logistic regression model was constructed using the above four indicators, and ROC showed AUC of this model for predicting LVI was 0.882, superior to any of the single indicator ( P<0.05); its sensitivity was 80.36% and specificity was 84.62%. Hosmer-lemeshow test showed that the prediction model had good calibration ( P=0.503). Conclusion:The combined prediction model constructed by preoperative breast MRI and axillary ultrasound could help to predict the LVI status of breast invasive ductal carcinoma.

The predictive model of microvascular invasion in hepatocellular carcinoma established based on multimodality imaging / 中华超声影像学杂志

Objective:To explore the risk factors of microvascular invasion (MVI) in hepatocellular carcinoma (HCC), and to predict MVI preoperatively, non-invasively and accurately.Methods:A total of 150 HCC patients (183 HCC lesions) were retrospectively collected in the First Affiliated Hospital of Xi′an Jiaotong University from January 2016 to June 2022.The clinical data and hematological data, gray-scale ultrasonography (US), contrast-enhanced ultrasonography (CEUS), enhanced magnetic resonance imaging with gadolinium ethoxybenzyl diethylenetriamine pentaacetic acid (EOB-MRI) and pathological data of these patients were recorded. According to the pathological diagnosis of MVI, the lesions were divided into MVI (+ ) group and MVI (-) group. The indicators between the two groups were compared. All 183 lesions were put into the training set, and the prediction model with nomogram was constructed according to the risk factors of MVI selected by multivariate Logistic regression. The internal verification was carried out by ten-fold cross-validation method.Results:There were significant statistical differences in the following parameters between MVI (+ ) group ( n=109) and MVI (-) group ( n=74) (all P<0.05). These were cirrhosis, serological parameters (alpha-fetoprotein, albumin, total bilirubin), qualitative indexes of US (size, boundary, internal echo), qualitative indexes of CEUS (hyper/iso/hypovascularity of lesions in arterial phase, portal phase, and delayed phase compared with hepatic parenchyma), and quantitative indexes of EOB-MRI [post enhancement rate (post ratio) and gadolinium disodium rate (EOB ratio)] calculated mainly in terms of lesions and surrounding liver parenchyma in hepatobiliary phase and unenhanced T1 images). Finally, cirrhosis of patients, the size, boundary, internal echo of lesions in US; arterial phase (AP), portal phase (PP), post-vascular phase (PVP) features in CEUS; the EOB rate and post rate of EOB-MRI entered the prediction model of MVI. The training set exhibited good calibration and net gain rate. The areas under the ROC curve for the training set and the validation set were 0.981 and 0.961, respectively, while the diagnostic accuracy were 92.9% and 85.8%, respectively. Conclusions:The model constructed mainly by multimodality imaging methods can achieve favorable predictive performance for MVI, which provides valuable ideas for noninvasively predicting the incidence of MVI and optimizing the MVI-related treatment of MVI in HCC patients.

Effect of transcription factor NFIB on cell proliferation and invasion in breast cancer / 中国医师杂志

Objective:To investigate the effect of transcription factor nuclear factor IB (NFIB) on cell proliferation and invasion in breast cancer.Methods:The lentivirus pLKO.1-shNFIB plasmid was constructed, packaged and infected with human estrogen receptor positive (ER + ) breast cancer cell line MCF-7 and triple-negative breast cancer (TNBC) cell line MDA-MB-231, respectively, NFIB was stably knocked down and verified by Western blot method; Cell count test (CCK-8) and clone formation test were used to investigate the effect of knockdown NFIB on the growth and proliferation of breast cancer cells; The transwell experiment and Western blot method were performed to detect the expression of epithelial mesenchymal transition protein markers. The effect of knockdown NFIB on the invasive ability of triple-negative breast cancer cells was explored; Kaplan-Meier survival was used to analyze web data (http: //kmplot.com/analysis/) to explore the effect of NFIB on the prognosis of ER + breast cancer and triple-negative breast cancer patients. Results:In MCF-7 and MDA-MB-231 breast cancer cells, knocking down NFIB inhibited cell growth and proliferation; In triple-negative breast cancer MDA-MB-231 cells, knocking down NFIB promoted the expression of interstitial marker fibronectin and promoted cell invasion; The lower the expression of NFIB, the worse the prognosis of triple negative breast cancer patients, while the expression of NFIB had no effect on the prognosis of ER + breast cancer patients. Conclusions:Knocking down NFIB inhibits the proliferation of MCF-7 cells, and the expression level of NFIB is not related to the prognosis of ER + breast cancer patients; Knocking down NFIB inhibits the proliferation of MDA-MB-231 cells but promotes their invasion; The low expression of NFIB is associated with the poor prognosis of triple-negative breast cancer patients.

Advances in preoperative imaging evaluation of extramural vascular invasion of rectal cancer / 中国医师杂志

Rectal cancer is a common malignancy of the gastrointestinal tract. Extramural vascular invasion (EMVI) is a key indicator of risk stratification for rectal cancer and an important reference factor in determining individualised treatment options, so it is important to accurately assess whether extramural vessels are infiltrated by the tumour before surgery. The main imaging methods for rectal cancer include magnetic resonance imaging (MRI), computed tomography (CT) and transrectal ultrasound (TRUS). This article focuses on the performance and diagnostic efficacy of these imaging methods in the diagnosis of EMVI in rectal cancer, and provides clinical reference.

Mechanism of berberine inhibiting invasion and migration of renal carcinoma cells by down-regulating the expression of METTL3 / 中国药房

OBJECTIVE To investigate the inhibitory effect of berberine （BER） on the invasion and migration of human renal carcinoma cells and its potential mechanism. METHODS Using human renal carcinoma OSRC-2 cell as object， alamarBlue assay was adopted to detect the inhibitory effects of 0 （control group）， 25， 50， 75， 100， 125， 150， 175 and 200 μmol/L BER on the proliferation of OSRC-2 cell after treatment for 24 h and 48 h. After treated with 0（control group）， 50， 100 μmol/L BER for 48 h， the effect of BER on cell cycle was analyzed by flow cytometry. The migration of OSRC-2 cells in 24 h and 36 h was observed by cell scratch test， and the invasion ability of OSRC-2 cells in 24 h was detected by Transwell assay. The protein expression of methyltransferase-like 3 （METTL3） was detected by Western blot after treatment for 48 h， and RNA methylation quantification kit was used to detect the levels of N6-methyladenosine （m6A） in OSRC-2 cells. RESULTS Compared with control group， BER at different concentrations could significantly decrease the survival rate of OSRC-2 cells （P＜0.01）， and showed a dose-dependent and time-dependent manner. After 48 h of BER treatment at 50， 100 μmol/L， the cell was arrested in G0/G1 phase （P＜0.01）. Compared with control group， the migration and invasion abilities of cells in 50， 100 μmol/L BER group were significantly decreased （P＜0.05 or P＜0.01）； the protein expression of METTL3 and the level of m6A in RNA were significantly decreased （P＜0.01）. CONCLUSIONS BER can inhibit level of m6A by down-regulating the expression of METTL3， thereby inhibiting the invasion and metastasis of human renal carcinoma cells.

Effects of β-sitosterol on the function of synovial fibroblasts in rheumatoid arthritis and its mechanism / 中国药房

OBJECTIVE To investigate the effects of β-sitosterol on the function of rheumatoid arthritis （RA） fibroblastic synoviocytes MH7A cells and its mechanism. METHODS Network pharmacology was adopted to screen the targets of β-sitosterol and the targets for the treatment of RA. After the intersection of them， topological analysis was performed to find the most critical target in the treatment of RA. MH7A cells were treated with different concentrations （0， 5， 10， 20， 40 μmol/L） of β-sitosterol， and CCK-8 was used to assay cell viability for screening the optimal concentration of β-sitosterol. MH7A cells were induced by 10 ng/mL TNF-α in vitro and treated with β-sitosterol （the optimum concentration）. CCK-8 and EdU were used to detect the ability of cell proliferation. Scratch experiment and Transwell invasion assay were used to analyze cell migration and invasion. The levels of interleukin-1β （IL-1β） and IL-6 in cell supernatant were detected by enzyme-linked immunosorbent assay （ELISA）. The mRNA and protein expressions of peroxisome proliferator-activated receptor α （PPARα） were measured by qRT-PCR and Western blot， respectively. The siRNA targeting PPARα was transfected into MH7A cells， and the effects of β-sitosterol on cell proliferation， migration， invasion， the secretion of inflammatory factors and the expression of PPARα after PPARα knockdown were detected by the above experimental methods. RESULTS PPARα was the most critical target of β-sitosterol in the treatment of RA. The optimal concentration of β-sitosterol was 20 μmol/L. Compared with model group， β-sitosterol decreased the viability of MH7A cells， and the number of proliferating cells also decreased significantly （P＜0.05）； the cell migration rate and the number of cell invasion decreased significantly （P＜0.05）. The levels of IL-1β and IL-6 were also significantly decreased （P＜0.05）， and the mRNA 15 and protein expression levels of PPARα were significantly increased （P＜0.05）. Compared with negative control small interfering RNA group， after PPARα knockdown， the cell viability increased by about 35.6% （P＜0.05）， the number of cell proliferation， the cell migration rate and the number of cell invasion increased significantly （P＜0.05）， and the levels of IL-1β and IL-6 also increased significantly （P＜0.05）. CONCLUSIONS β-sitosterol could effectively inhibit the proliferation， migration， invasion and secretion of inflammatory factors in MH7A cells， the mechanism of which may be associated with activating PPARα pathway.

Effects of Forsythia suspensa ethanol extract on the proliferation，migration and invasion of lung cancer cells NCI-H226 / 中国药房

OBJECTIVE To investigate the effects of Forsythia suspensa ethanol extract on the proliferation， migration and invasion of lung cancer cells NCI-H226. METHODS As research objects， lung cancer cells NCI-H226 were divided into control group， F. suspensa ethanol extract low-， medium- and high-concentration groups （5， 10， 20 mg/mL）， activator group [10 mg/mL F. suspensa ethanol extract+0.5 μmol/L nuclear factor kappa B （NF-κB） signaling pathway activator PMA]， inhibitor group （10 mg/mL F. suspensa ethanol extract+10 μmol/L NF-κB signaling pathway inhibitor BAY 11-7082） and positive control group （20 μg/mL cisplatin）. Except for the control group of cells without intervention， all other groups of cells were cultured with corresponding drugs for 24 hours； the proliferation， migration and invasion of cells were all detected， and the proliferation rate， migration rate， and the number of invading cells were also calculated； protein expressions of NF-κB p65， NF-κB inhibitory protein α （IκBα）， phosphorylated NF-κB p65 （p-NF-κB p65） and phosphorylated IκBα （p-IκBα） were determined. RESULTS Compared with control group， the proliferation rate， migration rate， and the number of invading cells as well as the protein expressions of p- IκBα and p-NF-κB p65 were decreased significantly in F. suspensa ethanol extract groups and positive control group （P＜0.05）. Compared with F. suspensa ethanol extract medium-concentration group， the proliferation rate， migration rate， and the number of invading cells as well as above protein expressions were all decreased significantly in inhibitor group （P＜0.05）， while those of activator group were increased significantly （P＜0.05）. CONCLUSIONS F. suspensa ethanol extract can inhibit the proliferation， migration and invasion of lung cancer cells NCI-H226， and the mechanism of which may be related to the inhibition of NF-κB signaling pathway.

Inhibitory effects of ursolic acid on IL-6-mediated invasion and migration of breast cancer cells / 中国药房

OBJECTIVE To investigate the inhibitory effects of ursolic acid on interleukin-6 （IL-6）-mediated invasion and migration of breast cancer MDA-MB-231 cells （hereinafter referred to as “231 cells”）. METHODS The effects of 20， 40， 80， 160 and 320 µmol/L ursolic acid on the proliferation rate of 231 cells were measured by CCK-8 method. The breast cancer 231 cells were divided into control group， model group and administration group. The migration and invasion abilities of cells were detected by scratch assay and Transwell assay. Real-time quantitative polymerase chain reaction （q-PCR） assay and Western blot assay were used to detect the mRNA and protein expressions of epithelial-mesenchymal transition-related makers such as E cadherin （E-cad）， matrix metalloprotein 2 （MMP2）， MMP9， vimentin （Vim）， CD44 molecule （CD44） and aldehyde dehydrogenase 1 family member A1 （ALDH1A1）. The phosphorylation levels of JAK2 and STAT3 in the Janus kinase 2/signal transducer and activator of transcription 3 （JAK2/STAT3） signaling pathway （in terms of p-JAK2/JAK2 ratio and p-STAT3/STAT3 ratio） were detected by Western blot assay. RESULTS A low concentration of ursolic acid of 20 µmol/L （no significant inhibitory effect on cell proliferation ability） was selected as the subsequent administration concentration. Compared with the control group， the migration and invasion abilities of cells in the model group were significantly enhanced （P＜0.05）； compared with the model group， the migration and invasion abilities of cells in the administered group were significantly reduced （P＜0.05）. Compared with the control group， the relative mRNA and protein expressions of epithelial-mesenchymal transition-related markers MMP9， MMP2， Vim， ALDH1A1 and CD44 were all elevated to different extents， and the mRNA and protein expressions of E-cad were all decreased to different extents in the model group cells， and part of the differences had statistical significance （P＜0.05）， the p-JAK2/JAK2 ratio and p-STAT3/STAT3 ratio were significantly increased in the model group （P＜0.05）； compared with the model group， the expressions of the above indicators were reversed to some extent in the administration group. CONCLUSIONS Ursolic acid blocks the activation of JAK2/STAT3 signaling pathwby the inflammatory factor IL-6， which ultimately interrupts the invasion and metastasis of breast cancer cells.

Overexpression of CLEC5A inhibits cell proliferation and metastasis and reverses epithelial-mesenchymal transition in hepatocellular carcinoma / 南方医科大学学报

OBJECTIVE@#To evaluate the effects of CLEC5A expression level on cell proliferation, migration and invasion and epithelial-mesenchymal transition (EMT) in hepatocellular carcinoma (HCC) and explore the role of CLEC5A in the tumorigenesis and progression of HCC.@*METHODS@#The expression level of CLEC5A was detected in 50 pairs of HCC and adjacent tissues using immunohistochemical staining, and its association with clinicopathological parameters of HCC patients was analyzed. Cultured HCC cell line SK-HEP-1 was transfected with a lentiviral vector overexpressing CLEC5A, and the transfection efficiency was verified using real-time fluorescence quantitative PCR and Western blotting. The changes in proliferation, migration and invasion abilities of the transfected cells were analyzed using CCK-8, 5-ethynyl-29-deoxyuridine (EdU) and Transwell assays, and EMT of the cells was determined using Western blotting.@*RESULTS@#The protein expression level of CLEC5A was significantly lower in HCC tissues than in the adjacent tissues (P < 0.001). The expression level of CLEC5A was significantly correlated with tumor size (P=0.008), tumor number (P=0.010), histological differentiation (P=0.016), microvascular invasion (P=0.024) and BCLC stage (P=0.040). In SK-HEP-1 cells, overexpression of CLEC5A obviously inhibited the cell proliferation, migration and invasion and reversed EMT phenotype of the cells.@*CONCLUSION@#CLEC5A is a potential HCC suppressor gene and may serve as a promising therapeutic target for HCC.