Value of detection of peripheral blood miR-194 combined with fecal miR-143 in the clinical screening of colorectal cancer / 国际肿瘤学杂志

Objective:To explore and analyze the value of detection of peripheral blood miR-194 combined with fecal miR-143 in the clinical screening of colorectal cancer.Methods:A total of 83 patients diagnosed with colorectal cancer by pathological tissue admitted to Huangshi Hospital of Traditional Chinese Medicine of Hubei Province from October 2019 to October 2020 were selected as the observation group, and 50 healthy volunteers who underwent physical examinations during the same period were selected as the control group. The levels of miR-194 in peripheral blood and miR-143 in feces were detected by fluorescence quantitative PCR. The level difference between the two groups and their correlations with clinicopathological parameters of patients with colorectal cancer were analyzed. Receiver operating characteristic (ROC) curve was drawn based on peripheral blood miR-194 and fecal miR-143 to evaluate their value for clinical screening of colorectal cancer.Results:The level of miR-194 in peripheral blood of the observation group was significantly higher than that of the control group (1.91±0.34 vs. 0.76±0.23) , while the level of fecal miR-143 in the observation group being significantly lower than that of the control group (1.85±0.43 vs. 2.48±0.62) , with statistically significant differences ( t=21.16, P<0.001; t=6.91, P<0.001) . Age of patients with colorectal cancer ( t=0.83, P=0.408; t=1.17, P=0.244) , TNM stage ( t=1.03, P=0.307; t=0.11, P=0.909) , lymphatic metastasis ( t=0.37, P=0.711; t=1.85, P=0.068) , distant metastasis ( t=0.41, P=0.683; t=1.72, P=0.089) were not correlated with the levels of peripheral blood miR-194 and fecal miR-143. When the cut-off value of miR-194 in peripheral blood was 1.82, the area under the ROC curve for the diagnosis of colorectal cancer was 0.76, and the diagnostic sensitivity and specificity were 79.38% and 74.29%, respectively. When the cut-off value of fecal miR-143 was 2.16, the area under the ROC curve for the diagnosis of colorectal cancer was 0.71. At this time, the diagnostic sensitivity and specificity were 76.54% and 73.61%, respectively. The area under ROC curve of combined detection for colorectal cancer was 0.81, and the diagnostic sensitivity and specificity were 83.46% and 75.43%, respectively. Conclusion:Peripheral blood miR-194 is highly expressed in colorectal cancer patients, and fecal miR-143 is low in colorectal cancer patients. The combined detection of the two has a high sensitivity for early diagnosis of colorectal cancer, which can provide important reference basis for early diagnosis of colorectal cancer and has high clinical application value.

The value of circulating miR-143 level in predicting early response to concurrent chemoradiotherapy in cervical cancer patients / 中华放射肿瘤学杂志

Objective:To investigate the value of serum miR-143 level combined with MRI in predicting the early response to concurrent chemoradiotherapy (CCRT) in cervical cancer.Methods:A total of 85 patients with pathologically confirmed cervical cancer underwent conventional MRI, intravoxel incoherent motion diffusion-weighted imaging (IVIM-DWI), and dynamic contrast-enhanced MRI (DCE-MRI) before CCRT. The biopsy tissues and serum samples were collected. The differential expression of miRNA in the biopsy tissues was determined by microarray chip. The expression level of miR-143 in the serum samples was analyzed by qRT-PCR. All patients were divided into the non-residual and residual tumor groups according to post-treatment MRI. Pre-treatment clinical factors, MRI parameters and miR-143 between two groups were statistically analyzed by the univariate and multivariate analyses. The optimal thresholds and predictive performance for post-treatment incidence of residual tumors were estimated by drawing the ROC curve.Results:At one month after CCRT, there were 52 patients in the non-residual tumor group and 33 patients in the residual tumor group. In the residual tumor group, pre-treatment FIGO staging, apparent diffusion coefficient (ADC), D and V e were significantly higher (all P<0.05), whereas K trans value was significantly lower ( P<0.001) when compared to those in the non-residual tumor group. The miRNA array analysis showed that there were 16 miRNAs with differential expression levels between two groups (all P<0.05). Among them, the increase of miR-143 was the most significant in the residual tumor group. Compared with the residual tumor group, the expression level of serum miR-143 was significantly down-regulated in the non-residual tumor group ( P=0.002). Compared with the SiHa cells, the expression level of miR-143 in the SiHa-R cells was significantly up-regulated ( P<0.05). Multivariate analysis showed that only miR-143, D, K trans and V e were the independent prognostic factors. The combination of multi-parametric MRI and miR-143 exhibited the highest predictive performance (AUC=0.975), with a sensitivity of 84.8% and a specificity of 96.2%. Conclusion:The combination of multi-parametric MRI with miR-143 further improves the predictive performance for residual tumors after CCRT, which contributes to the personalized treatment of cervical cancer.

lncRNA OIP5-AS1 targets ROCK1 to promote cell proliferation and inhibit cell apoptosis through a mechanism involving miR-143-3p in cervical cancer

Opa-interacting protein 5 antisense transcript 1 (OIP5-AS1) is one kind of cytoplasmic long non-coding RNA (lncRNA), which has been demonstrated to play a critical function in multiple cancers. However, the detailed mechanism of OIP5-AS1 in the regulation of cervical cancer progression is still obscure. Here, we demonstrated that lncRNA OIP5-AS1 was upregulated in cervical cancer and was correlated with poor prognosis by bioinformatics studies. OIP5-AS1 depletion inhibited cell proliferation and promoted cell apoptosis in cervical cancer cells. Furthermore, we clarified that ROCK1 was the downstream effector of OIP5-AS1 and OIP5-AS1 acted as a molecular sponge of miR-143-3p. Finally, we verified that OIP5-AS1 exerted its function in the regulation of cervical cancer progression via interacting with miR-143-3p to regulate ROCK1 expression. Our study revealed novel mechanisms about how lncRNA OIP5-AS1 executed its function in cervical cancer and thus provided potential therapeutic targets for the disease.

Clinical significance of serum miR-187 and miR-143 in the diagnosis of gallbladder cancer / 中华肝胆外科杂志

Objective To investigate the clinical significance of serum miR-187 and miR-143 in the development and diagnosis of gallbladder cancer.Methods 75 serum samples in patients with gallbladder cancer were selected as gallbladder cancer group.75 serum samples in patients with gallbladder benign disease and 45 serum samples in healthy physical examinations at same period were selected as the benign gallbladder disease group and healthy control group.Quantitative RT-PCR was used to detect the serum miR-187 and miR-143 expression in each group,and the expression of those related with the clinicopathological factors,the proliferation and migration of gallbladder cancer cells,and the efficacy in diagnosis of gallbladder cancer was observed.Results The serum miR-187 expression in gallbladder cancer group was significantly higher than that in benign gallbladder disease and healthy control;the serum expression of that in benign gallbladder disease was significantly higher than that in healthy control;after surgery,the expression of that was significantly lower than that before treatment (all P ＜ 0.05).The serum expression of miR-143 in gallbladder cancer group was significantly lower than that in benign gallbladder disease and healthy control;the serum expression of that in benign gallbladder disease was significantly lower than that in healthy control;after surgery the serum expression of that was significantly higher than that before surgery (all P ＜0.05).The serum expression levels of miR-187 and miR-143 in gallbladder cancer were not correlated with gender and age (both P ＞ 0.05),and were significantly correlated with Nevin stage,TNM stage,differentiation and lymphatic metastasis (all P ＜ 0.05).Furthermore,it was confirmed that miR-187 promoted the proliferation and migration of gallbladder cancer cells in vitro,while miR-143 inhibited the proliferation and migration.In the diagnosis of gallbladder,the diagnostic efficacy of miR-187 and miR-143 was significantly better than that of CA199 and CA242 (both P ＜ 0.05).Combined detection could further improve the efficacy in diagnosis of gallbladder cancer.Conclusions miR-187 and miR-143 are involved in the development of gallbladder cancer.Combined detection of serum miR-187 and miR-143 in gallbladder cancer has a high diagnostic efficiency in the diagnosis of gallbladder cancer.

miR-143-3p regulates proliferation, migration and invasion of colon cancer RKO cells via targeting EZH2 / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the molecular mechanism of miR-143-3p regulating the proliferation, migration and invasion of colon cancer RKO cells via targeting enhancer of zeste homolog 2 (EZH2). Methods: A total of 40 pairs of colon cancer tissues and corresponding para-cancerous tissues resected in the First Affiliated Hospital of Kunming Medical University from March 2015 to July 2017 were collected for this study. In addition, colon cancer cell lines (COLO320, RKO and CL-11) and normal intestinal mucosa NCM460 cells were also collected. qPCR was applied to detect the expression level of miR-143-3p in colon cancer tissues and cell lines. miR-143-3p mimics, miR-143-3p inhibitor, EZH2 siRNA and negative control plasmids were transfected into RKO cells, respectively. The effect of miR-143-3p/EZH2 axis on the proliferation, migration and invasion of RKO cells were detected by CCK-8 and Transwell assay, respectively. Western blotting was used to detect the expression level of EZH2 protein in RKO cells. The targeting relationship between miR-143-3p and EZH2 was verified by Dual luciferase reporter gene assay. Results: The expression level of miR-143-3p was downregulated in colon cancer tissues and cell lines (all P<0.01). Overexpression of miR-143-3p significantly inhibited the proliferation, migration and invasion of RKO cells (all P<0.01). Dual luciferase reporter gene assay confirmed that EZH2 was a target gene of miR-143-3p. Simultaneous knockdown of miR-143-3p and EZH2 attenuated the inhibition of EZH2 knockdown on the proliferation, migration and invasion of RKO cells. Conclusion: miR-143-3p suppresses the proliferation, migration and invasion of colon cancer cells via targetedly down-regulating EZH2.

Effects of miR-143 target-regulating ER-α36 on the invasion of gastric cancer cell line SGC 7901 / 基础医学与临床

Objective To investigate the effects of ER-α36 on invasion of human gastric cancer cell lines SGC7901 by miR-143. Methods Lentiviral vectors were constructed to generate up-and down-regulations of miR-143 lentiviruses (LV-miR-143 and LV-anti-miR-143,respectively).The viruses were used to infect human gastric cancer cell lines SGC7901.The ER-α36 protein expression level and the invasion of constructed cells were detected by Western blot and transwell. The target gene of miR-143 was predicted by bioinformatics tools.Luciferase reporter assay was carried out to confirm the predicted target gene. Results The infection efficiency of the lentivirus titers of LV-miR-143 and LV-anti-miR-143 were over 80% shown by the green fluorescence. The ER-α36 expression level,the cell invasion in LV-miR-21 group were significantly lower than those in LV-anti-miR-21 group(P<0.05). Conclusions miR-143 plays an important role in the negative control of gastric cancer invasion by the regulation of ER-α36.

lncRNA CCAT1 promotes cell proliferation, migration, and invasion by down-regulation of miR-143 in FTC-133 thyroid carcinoma cell line

Thyroid cancer is a common malignant tumor. Long non-coding RNA colon cancer-associated transcript 1 (lncRNA CCAT1) is highly expressed in many cancers; however, the molecular mechanism of CCAT1 in thyroid cancer remains unclear. Hence, this study aimed to investigate the effect of CCAT1 on human thyroid cancer cell line FTC-133. FTC-133 cells were transfected with CCAT1 expressing vector, CCAT1 shRNA, miR-143 mimic, and miR-143 inhibitor, respectively. After different treatments, cell viability, proliferation, migration, invasion, and apoptosis were measured. Moreover, the regulatory relationship of CCAT1 and miR-143, as well as miR-143 and VEGF were tested using dual-luciferase reporter assay. The relative expressions of CCAT1, miR-143, and VEGF were tested by qRT-PCR. The expressions of apoptosis-related factors and corresponding proteins in PI3K/AKT and MAPK pathways were analyzed using western blot analysis. The results suggested that CCAT1 was up-regulated in the FTC-133 cells. CCAT1 suppression decreased FTC-133 cell viability, proliferation, migration, invasion, and miR-143 expression, while it increased apoptosis and VEGF expression. CCAT1 might act as a competing endogenous RNA (ceRNA) for miR-143. Moreover, CCAT1 activated PI3K/AKT and MAPK signaling pathways through inhibition of miR-143. This study demonstrated that CCAT1 exhibited pro-proliferative and pro-metastasis functions on FTC-133 cells and activated PI3K/AKT and MAPK signaling pathways via down-regulation of miR-143. These findings will provide a possible target for clinical treatment of thyroid cancer.

Differential expression of miR-143 in colon cancer tissue and its effect on apoptosis and migration of colon cancer cell / 上海交通大学学报（医学版）

Objective · To study the expression of microRNA-143 (miR-143) in human colon cancer and adjacent normal tissues, and to explore its effect on the function of colon cancer cells and downstream target genes. Methods · Lentiviral expression vector of miR-143 was designed and used to stably transfect colon cancer cell lines HCT116 and RKO. Real-time PCR was used to detect the expression of miR-143 in colon cancer, adjacent normal tissues, HCT116 cells and RKO cells infected with lentivirus. The apoptosis ratio of HCT116 cells and RKO cells was detected by Annexin V-APC single staining and flow cytometry. The migration ability of HCT116 cells and RKO cells was detected by Transwell method. The expression of FOSL2 protein was detected by Western blotting assay. Results · The expression of miR-143 in colon cancer tissue was significantly lower than that in adjacent mucosal tissue (0.339±0.454 vs 1.003±0.003) (U=16.000, Z=-4.231, P=0.000). By up-regulation of miR-143, apoptosis rates were significantly higher in HCT116 and RKO cells than those in the negative controls (P=0.000). The count of migrating HCT116 cells in up-regulated group was significantly lower than that in the negative control group (P=0.000). The count of migrating RKO cells in down-regulated group was significantly higher than that in the negative control (P=0.003). By down-regulation of miR-143, the expression of FOSL2 protein was increased. Conclusion · The expression of miR-143 was significantly decreased in colon cancer tissues. Up-regulation of miR-143 can promote the apoptosis rate in both cell lines, and inhibit the migration ability of HCT116 cells. Down-regulation of miR-143 could promote the migration ability of RKO cells. By inhibiting miR-143, the expression of FOSL2 protein in colonic cancer cells would be increased.

Effect of miRNA-143 on proliferation, migration and invasion of CNE-2Z cells of human nasopharyngeal cancer / 安徽医科大学学报

Objective To detect the effects of miR-143 on proliferation, migration and invasion of human nasopharyngeal cancer CNE-2Z cells.Methods The expression of miR-143 in NP69 cells, CNE-1 cells and CNE-2Z cells were detected by using real-time PCR.The overexpression of miR-143 in CNE-2Z cells was constructed through infecting lentivrius, and the level of miR-143 was determined by using real-time PCR.Cell viability was detected by using a CCK-8 kit according to the manufacturer's instruction at indicated time points.The effectiveness of overex-pression of miR-143 on migration and invasion of CNE-2Z cells were detected by using Transwell cell migration and invasion assay.Results The expression level of miR-143 in CNE-2Z was significantly lower than those in NP69 and CNE-1 (P<0.01).The miR-143 over-expressed CNE-2Z cell was successfully established.The cell viability in CNE-2Z/ miR-143 group was significantly decreased compared with CNE-2Z and CNE-2Z/miR-NC (P<0.01).Overexpression of miR-143 inhibited cell migration and invasion of CNE-2Z cells significantly(P<0.01).Conclusion miR-143 might inhibit cell proliferation, migration and invasion in human nasopharyngeal cancer CNE-2Z cells, indicating its important role in diagnosis, treatment and prognosis of nasopharyngeal cancer.

Effects of miR-143 on invasion and migration of B cell lymphoma cells through regulating expression of PAN3 protein / 中国免疫学杂志

Objective:To investigate the effect of miR-143 on the invasion and migration of B cell lymphoma cells .Methods:The expression of miR-143 in normal bone marrow and lymphoma was detected by qPCR .The expression levels of miR-143 in different cell lines were examined by qPCR .qPCR was used to detect the ability of miR-143 on PAN3.The relationship between miR-143 and PAN3 was detected by double luciferase assay .The effect of miR-143 expression on the migration ability of B cell lymphoma E 6-1 cells was examined by scratch test .The effect of miR-143 expression on the invasion ability of B cell lymphoma E 6-1 cells was exa mined by transwell test.Results:Compared with normal bone marrow ,miR-143 was down-regulated in B-cell lymphoma.Double luciferase assay showed that miR-143 could regulate the expression of PAN 3.Overexpression of miR-143 ,E6-1 cells significantly reduced the ability to attack and migrate.Conclusion:miR-143 can regulate the migration and invasion of B cell lymphoma cells by regulating the expression of PAN3.

Expression of miR-143 in patients with PTC and its influence on immune function / 中国免疫学杂志

Objective:To investigate the relationship between the expression level of miR 143 and prognosis/cellular biological characteristics in patients with papillary thyroid carcinoma (PTC),and explore the potential mechanisms.Methods:30 PTC patients were admitted and enrolled into the trial from 2013 April to 2016 December.The qPCR was performed to detect the expression of miR143 in cancer tissues and adjacent tissues from 30 PTC patients.The effect of miR143 on overall survival rate and metastasis-free survival rate was assessed with specific analysis .CON and pGenesil1-miR143 vector were transfected into PTC cell line IHH-448h after qPCR.CKK8 and Annexin V/PI flow cytometry were performed to detect cell proliferation ,and apoptosis,respectively.Cell migration was asseessed by transwell experiment .Levels of miR143,IL-10,IFN-γin serum were assessed and calculated with the pearson coeffi-cient.PTC patients were assigned into miR 143 high and low expression group according to miR 143 expression , and IL-10 and IFN-γwere measured with routine protocol.Results:The relative expression of miR143 in paracancerous tissues and tumor tissues was (1.0± 0.15) vs (0.12±0.06).miR143 was significantly associated with the survival rate and metastasis rate of patients (P<0.001).miR143 expression levels in IHH-4 pGenesil1-miR143 and control group were ( 8.63 ±0.71 ) vs ( 1.0 ±0.06 ) , and the proliferative index in pGenesil1-miR143 group was significantly lower than CON group (P<0.01).Percentage of Annexin and V+positive cells in pGenesil1-miR143 and CON group was (40±7.2) vs (2±0.38),and the ratio of the number of migrating cells was (40±7.2) vs (2±0.38) (P<0.001).Analysis showed that r2 were -0.4 and 0.62 for IL-10 and IFN-γ,respectively(P<0.001).In miR143 low group and high ex-pression group,expression levels of IFN-γwas (8±0.23) vs (11.2±0.12),while IL-10 was (12±3.1) vs (8.43±0.44) (P<0.05). Conclusion:miR143 is low expressed in PTC patients and associated with prognosis and survival .Highly expressed miR143 inhibited the proliferation of IHH-4 cell lines,increased apoptosis and decreased migration ,also the expression of miR143 is related to the immune function of the patients .

miR-143 suppresses the proliferation and migration of SGC7901 gastric cancer cells / 中国肿瘤临床

Objective:To investigate the role and mechanism of miR-143 in the proliferation and migration of gastric cancer (GC) cells. Methods:Western blot was performed to detect the expression level of avian erythroblastosis oncogene B-3 (ERBB3) in GC tissues, paired non-cancerous tissues, and SGC7901 GC cells. RT-qPCR was conducted to determine the mRNA and miR-143 of ERBB3 quantita-tively. Bioinformatics tools were used to predict the target gene of miR-143. Luciferase reporter assay was carried out to confirm the predicted target gene. Transwell and EdU assays were applied to observe the migration and proliferation of SGC7901 GC cells transfect-ed with miR-143 mimics/inhibitor/NC mimics/inhibitor. Results:Compared with the expression levels of ERBB3 and miR-143 in the paired non-cancerous tissues, the expression level of ERBB3 was upregulated and the expression level of miR-143 was downregulated in GC tissues. In the prediction of the potential target gene, miR-143 could bind to a specific sequence of the 3′-untranslated regions (UTR) of the mRNA of ERBB3. This finding was supported by luciferase reporter assay results. In vitro, ERBB3 protein expression and cell migration and proliferation were suppressed significantly in the SGC7901 cells transfected with miR-143 mimics. By contrast, these processes were remarkably enhanced when the cells were transfected with miR-143 inhibitor. Conclusion:miR-143 can suppress the migration and proliferation of GC cells by downregulating the expression of ERBB3.