Cancer-associated fibroblast-secreted miR-421 promotes pancreatic cancer by regulating the SIRT3/H3K9Ac/HIF-1α axis.

This study was designed to explore the effects of exosomal miR-421 secreted by cancer-associated fibroblasts (CAFs) on pancreatic cancer (PC) progression and the mechanisms involved. CAFs and exosomes (exos) were isolated and identified. PC cells were treated with CAF-derived exos (CAF-exos). Western blotting and quantitative polymerase chain reaction (qPCR) were used to measure miR-421, sirtuin-3 (SIRT3), and hypoxia duciblefactors-1 alpha (HIF-1α) levels. Cell counting kit-8 (CCK-8), wound-healing, and transwell migration assays were used to measure proliferation, migration, and invasion abilities of the cells. Dual-luciferase assay and RNA immunoprecipitation (RIP) experiment analyzed the relationship between miR-421 and SIRT3. Chromatin immunoprecipitation (f)-verified H3K9Ac enrichment in the HIF-1α promoter region. In vivo tumorigenesis experiments were performed to further explore the effects of exosomal miR-421 from CAFs on PC. CAFs and exos were successfully isolated. CAF-exo-treated PC cells highly expressed miR-421 and had increased cell proliferation, migration, and invasion abilities. Knocking down miR-421 increased the expression of SIRT3. SIRT3 is a target of miR-421, and inhibiting the expression of SIRT3 reversed the negative effects of miR-421 knockdown on PC cell. Knocking down miR-421 in CAF-exo inhibited the expression of HIF-1α in PC cells. Moreover, SIRT3-mediated HIF-1α expression by regulating H3K9Ac. HIF-1α overexpression reversed the inhibiting effects of SIRT3 overexpression on PC progression and counteracted the inhibiting effects of miR-421 knockdown on glycolysis. Moreover, in vivo tumorigenesis experiments showed that knocking down miR-421 attenuated CAF-exo induced tumor growth. Exosomal miR-421 from CAFs promoted PC progression by regulating the SIRT3/H3K9Ac/HIF-1α axis. This study provided insights into the molecular mechanism of PC.

Development and validation of a nomogram to predict anastomotic leakage in colorectal cancer based on CT body composition.

Background: Anastomotic leakage (AL) is one of the most serious postoperative complications. This study aimed to investigate the predictive value of preoperative body composition for AL in patients with colorectal cancer (CRC). Methods: We first reviewed data from 3,681 patients who underwent radical CRC resection 2013-2021 in our hospital, and 60 patients were diagnosed with AL after surgery. We designed a nested case-control study and two controls were randomly selected for each case according to the time and position of surgery. Body composition was measured at the level of the third lumbar vertebra based on computed tomography (CT) images. The risk factors of AL were analyzed by univariate and multivariate analysis. Nomogram was built using binary regression analysis and assessed for clinical usefulness, calibration, and discrimination. Results: In the multivariate analysis, gender, blood glucose, nutrition risk screening (NRS), skeletal muscle area (SMA) and visceral fat area (VFA) were independent risk factors for developing anastomotic leakage after surgery. The prognostic model had an area under the receiver operating characteristic curve of 0.848 (95% CI, 0.781-0.914). The calibration curve showed good consistency between the predicted and observed outcomes. Decision curve analysis indicated that patients with colorectal cancer can benefit from the prediction model. Conclusions: The nomogram that combined with gender, blood glucose, NRS, SMA, and VFA had good predictive accuracy and reliability to AL. It may be conveniently for clinicians to predict AL preoperatively and be useful for guiding treatment decisions.

LINC00960 regulates cell proliferation and glycolysis in pancreatic cancer through the miR-326-3p/TUFT1/AKT-mTOR axis.

Pancreatic cancer (PC) is a common malignant cancer characterized by high mortality and poor prognosis. LINC00690 was involved in the occurrence and progression of PC, but the underlying mechanisms require further investigation. The goal of this study was to figure out how LINC00960 mediates glycolysis in PC. LINC00960, miR-326-3p, and Tuftelin 1 (TUFT1) expression levels were detected in PC cell lines. LINC00960 and TUFT1 expression levels were increased in PC cells when compared with normal pancreatic cells, whereas miR-326-3p expression levels were decreased. The expression levels of LINC00690 affected glycolysis in PC, and inhibition of LINC00960 inhibited tumor growth in vivo. LINC00690 targeted and suppressed the expression of miR-326-3p. MiR-326-3p bound to TUFT1, and miR-326-3p inhibited AKT-mTOR pathway activation via TUFT1. In conclusion, the depletion of LINC00960 repressed cell proliferation and glycolysis in PC by mediating the miR-326-3p/TUFT1/AKT-mTOR axis. Thus, we present a novel mechanism underlying the progression of PC that suggests LINC00960 is a potential therapeutic target for this cancer.

Detection and analysis of null alleles of amelogenin in gender identification.

In this study, we located eight samples with null alleles of amelogenin out of 10,750 cases, and discussed the influence in gender identification and forensic personal identification. Amelogenin was detected and retested by several autosomal STR kits and sex chromosomal STR kits, and the causes were analyzed by chromosome karyotype analysis and Y chromosome microdeletion detection if necessary. Suspected AMEL-X loss was observed in five samples, but no abnormality was detected in the X-STR loci. AMEL-X was recovered when samples were retested by other detection systems designed with different primers. One sample had AMEL-X and X-STR loci loss, and the karyotype was chimeric 45,X0[70]/46,X,+mar[13].Two male samples lost AMEL-Y fragment, and both of them lost DYS522-DYS570-DYS576 loci, but no abnormalities were found in the STS loci of SRY and AZF regions. Therefore, when carrying out gender identification by using amelogenin, it is essential to focus on null alleles of amelogenin. In especially, deal with the samples collected from the individuals who had chromosomal hereditary disorders(e.g. Turner Syndrome and Oligospermia / Azoospermia). In order to achieve this, laboratories should have various techniques to verify the null alleles of amelogenin and ensure accurate genotyping. Accurate genotyping of amelogenin and DNA database establishment are vital for personal identification.

Establishment and clinical application value of an automatic diagnosis platform for rectal cancer T-staging based on a deep neural network.

BACKGROUND: Colorectal cancer is harmful to the patient's life. The treatment of patients is determined by accurate preoperative staging. Magnetic resonance imaging (MRI) played an important role in the preoperative examination of patients with rectal cancer, and artificial intelligence (AI) in the learning of images made significant achievements in recent years. Introducing AI into MRI recognition, a stable platform for image recognition and judgment can be established in a short period. This study aimed to establish an automatic diagnostic platform for predicting preoperative T staging of rectal cancer through a deep neural network. METHODS: A total of 183 rectal cancer patients' data were collected retrospectively as research objects. Faster region-based convolutional neural networks (Faster R-CNN) were used to build the platform. And the platform was evaluated according to the receiver operating characteristic (ROC) curve. RESULTS: An automatic diagnosis platform for T staging of rectal cancer was established through the study of MRI. The areas under the ROC curve (AUC) were 0.99 in the horizontal plane, 0.97 in the sagittal plane, and 0.98 in the coronal plane. In the horizontal plane, the AUC of T1 stage was 1, AUC of T2 stage was 1, AUC of T3 stage was 1, AUC of T4 stage was 1. In the coronal plane, AUC of T1 stage was 0.96, AUC of T2 stage was 0.97, AUC of T3 stage was 0.97, AUC of T4 stage was 0.97. In the sagittal plane, AUC of T1 stage was 0.95, AUC of T2 stage was 0.99, AUC of T3 stage was 0.96, and AUC of T4 stage was 1.00. CONCLUSION: Faster R-CNN AI might be an effective and objective method to build the platform for predicting rectal cancer T-staging. TRIAL REGISTRATION: chictr.org.cn: ChiCTR1900023575; http://www.chictr.org.cn/showproj.aspx?proj=39665.

Establishment and application of an artificial intelligence diagnosis system for pancreatic cancer with a faster region-based convolutional neural network.

BACKGROUND: Early diagnosis and accurate staging are important to improve the cure rate and prognosis for pancreatic cancer. This study was performed to develop an automatic and accurate imaging processing technique system, allowing this system to read computed tomography (CT) images correctly and make diagnosis of pancreatic cancer faster. METHODS: The establishment of the artificial intelligence (AI) system for pancreatic cancer diagnosis based on sequential contrast-enhanced CT images were composed of two processes: training and verification. During training process, our study used all 4385 CT images from 238 pancreatic cancer patients in the database as the training data set. Additionally, we used VGG16, which was pre-trained in ImageNet and contained 13 convolutional layers and three fully connected layers, to initialize the feature extraction network. In the verification experiment, we used sequential clinical CT images from 238 pancreatic cancer patients as our experimental data and input these data into the faster region-based convolution network (Faster R-CNN) model that had completed training. Totally, 1699 images from 100 pancreatic cancer patients were included for clinical verification. RESULTS: A total of 338 patients with pancreatic cancer were included in the study. The clinical characteristics (sex, age, tumor location, differentiation grade, and tumor-node-metastasis stage) between the two training and verification groups were insignificant. The mean average precision was 0.7664, indicating a good training effect of the Faster R-CNN. Sequential contrast-enhanced CT images of 100 pancreatic cancer patients were used for clinical verification. The area under the receiver operating characteristic curve calculated according to the trapezoidal rule was 0.9632. It took approximately 0.2 s for the Faster R-CNN AI to automatically process one CT image, which is much faster than the time required for diagnosis by an imaging specialist. CONCLUSIONS: Faster R-CNN AI is an effective and objective method with high accuracy for the diagnosis of pancreatic cancer. TRIAL REGISTRATION: ChiCTR1800017542; http://www.chictr.org.cn.

Pancreatic stellate cells facilitate pancreatic cancer cell viability and invasion.

The biological features of pancreatic cancer and the associated hypoxic environment around the cancer cells often lead to resistance to radiotherapy and chemotherapy. The present study was performed in order to explore the effect pancreatic stellate cells (PSCs) have on the proliferation of pancreatic cancer cells. In the present study, PSCs from human pancreatic cancer tissues were isolated, and the PSCs markers α-smooth muscle actin and desmin were overexpressed in the cytoplasm of PSCs. An MTT assay revealed that PSCs promoted the viability of pancreatic cancer cells. However, the viability of pancreatic cancer cells promoted by PSCs was partially blocked by SB525334. Cellular invasion analysis demonstrated that PSCs promoted the invasion ability of pancreatic cancer cells. An apoptosis assay indicated that PSCs decreased the level of apoptosis induced by gemcitabine. In vivo experiments consisting of mice bearing MIA-PaCa-2 and PSCs demonstrated an increase in the rate of tumor growth compared with MIA-PaCA-2 alone, whereas SB525334 may delay the tumor progression induced by PSCs. The present findings indicated that PSCs promoted the viability and invasion of pancreatic cancer cells, and decreased the apoptosis of pancreatic cancer cells induced by gemcitabine.

Exosomes as critical mediators of cell-to-cell communication in cancer pathogenesis and their potential clinical application.

Exosomes are small membrane vesicles that measure 20 to 100 nm in diameter and are released by many cell types, including lymphocytes, dendritic cells (DCs) and tumor cells. As efficient messengers in cell-to-cell communication, exosomes released by tumors play an important role in regulating tumor malignancy. Tumor-derived exosomes contain proteins, mRNAs, and miRNAs, which can be delivered between different types of cells and even transferred to distant locations to influence the biological activities of tumors, such as proliferation, invasion and metastasis, immunoregulation, generation of a premetastatic niche and stimulation of angiogenesis. This review highlights advances in the understanding of exosome secretion and the role of exosomes in cancer molecular behavior. Moreover, we also discuss the potential clinical application of exosomes as biomarkers and therapeutic tools. Tumor-derived exosomes may represent a target for therapeutic intervention and for the development of early diagnostic biomarkers.

Pancreatic stellate cells increase pancreatic cancer cells invasion through the hepatocyte growth factor /c-Met/survivin regulated by P53/P21.

Pancreatic stellate cells (PSCs) are a key cellular component of the pancreatic tumor microenvironment and are considered to contribute to tumor invasion and metastasis. Multiple cytokines and growth factors derived from PSCs are involved in malignant cancer progression, including hepatocyte growth factor (HGF). However, the molecular mechanisms by which HGF regulates cancer invasion and metastasis have not been completely elucidated. Here, we report that two pancreatic cancer (PC) cell lines, Panc-1 and SW1990, displayed different invasive and migratory abilities after treatment with HGF secreted by PSCs. We found that HGF enhanced the invasive and migratory capacity of Panc-1 cells because of P53 deficiency, leading to overexpression of c-Met, which was regulated through P21. Additionally, our data showed that HGF/c-Met-mediated invasion and migration required the upregulation of survivin expression. In conclusion, PSCs promote PC cells invasion and migration via the HGF/c-Met/survivin pathway, which is negatively regulated by P53/P21.

Splenic Preservation Versus Splenectomy During Distal Pancreatectomy: A Systematic Review and Meta-analysis.

BACKGROUND: Studies have been published comparing spleen-preserving distal pancreatectomy (SPDP) with distal pancreatectomy with splenectomy (DPS), but the results remain inconsistent. The aim of this study was to compare SPDP with DPS by conducting a systematic review and meta-analysis. METHODS: Literature searches of the Medline/PubMed, Embase, and Cochrane Library databases were performed to identify relevant studies published before April 30,2015. Perioperative outcomes of SPDP and DPS were evaluated. The meta-analysis was performed in random- or fixed-effects models, as appropriate. A subanalysis was conducted to compare the two techniques of splenic preservation: splenic vessel preservation (SVP) and Warshaw technique (WT). RESULTS: Eighteen studies and 1156 patients were included in the comparison between SPDP and DPS. A total of 502 of these patients underwent SPDP and 654 underwent DPS. Meta-analysis showed the SPDP group had significantly fewer infectious complications (odds ratio [OR] 0.57, P = 0.006), less operative blood loss (P<0.0001), lower overall morbidity rate (OR 0.66, P = 0.002), and lower clinical pancreatic fistula rate (OR 0.42, P = 0.002) than the DPS group. Subanalysis indicated the SVP group had significantly lower rate of spleen infarction (OR 0.12, P<0.00001) and fewer secondary splenectomies (OR 0.13, P = 0.008) than the WT group. CONCLUSIONS: SPDP was a safe procedure associated with better short-term outcomes than DPS. SVP could provide more sufficient blood perfusion for the conserved spleen than WT. However, the evidence is limited, and more randomized controlled trials are warranted.

Diagnostic accuracy of K-ras mutation for pancreatic carcinoma: a meta-analysis.

BACKGROUND: The conventional tests for the diagnosis of early stage pancreatic carcinoma are not acceptable. This meta-analysis is to evaluate the accuracy of K-ras mutation for the diagnosis of pancreatic carcinoma. DATA SOURCES: A systemic search of all relevant literature was performed in Web of Science, EMBASE, Cochrane Database, and MEDLINE (PubMed as the search engine) prior to June 1, 2011. Thirty-four studies fulfilled the inclusion criteria and data were pooled for analysis. RESULTS: The pooled estimates for K-ras mutation in diagnosis of pancreatic carcinoma were as follows: sensitivity 0.68 (95% CI: 0.66-0.71), specificity 0.87 (95% CI: 0.85-0.88), positive likelihood ratio 4.54 (95% CI: 3.47-5.94), negative likelihood ratio 0.37 (95% CI: 0.30-0.44) and diagnostic odds ratio 14.90 (95% CI: 10.02-22.15). Summary receiver operating characteristic analysis demonstrated that the maximum joint sensitivity and specificity was 0.79, and the overall area under the curve was 0.86. CONCLUSIONS: Diagnostic accuracy of K-ras mutation was not superior to that of conventional tests. Therefore, K-ras mutation analysis alone is not recommended for the diagnosis of pancreatic carcinoma.

Vitamin D status and the risk of pancreatic cancer: a meta-analysis.

BACKGROUND: Vitamin D status in relation to pancreatic cancer risks is still inconsistent. This study was performed to evaluate the association between vitamin D status and risk of pancreatic cancer using a meta-analysis approach. METHODS: A systemic review of all relevant literature in English was performed by searching Pubmed, Web of Science and Embase to identify eligible studies from the earliest available date to April 1, 2012. The search terms "vitamin D", "25-hydroxyvitamin D", "pancreatic cancer" or "pancreatic neoplasms" were used to retrieve relevant papers. Inclusion criteria were: (1) the exposure of interest was intake of vitamin D or blood levels of vitamin D; (2) the outcome of interest was pancreatic cancer; (3) data on high and low intake or blood vitamin D in cases and controls were available; (4) odds ratio (OR) estimates with 95% confidence interval (CI) were provided; (5) primary epidemiological data were provided reporting pancreatic cancer incidence. The combined OR values and their 95% CIs were calculated via a meta-analysis. The potential presence of publication bias was estimated using Egger's regression asymmetry test. RESULTS: Nine studies with a total of 1 206 011 participants met the inclusion criteria. The test for heterogeneity showed there were significant differences among the included studies (I(2)=70.9%, P=0.001), so a randomized-effects model was used in the meta-analysis. The pooled OR of pancreatic cancer for the highest versus the lowest categories of vitamin D level was 1.14 (95% CI 0.896-1.451), and the Z-score for the overall effect was 1.06 (P=0.288), showing that there was no significant association between vitamin D levels and the risk of pancreatic cancer. Egger's test indicated there was a low possibility of publication bias in this study (P=0.348). CONCLUSION: Dietary vitamin D or circulating concentrations of 25-hydroxyvitamin D are not associated with the risk of pancreatic cancer based on evidence from currently published studies.

Optimized dose of imatinib for treatment of gastrointestinal stromal tumors: a meta-analysis.

OBJECTIVE: To investigate the efficacy and safety of standard-dose versus high-dose imatinib. METHODS: After a systematic review of English language articles, five studies including 2008 patients were eligible for the meta-analysis. Data extracted from each study were synthesized into overall odds ratios (OR). RESULTS: The overall OR for the high dose vs standard dose was 1.19 (95% CI 1.00-1.42) and the Z-score for the overall effect was 1.93 (P = 0.054), suggesting that high-dose imatinib added no survival benefits. The dose-related toxicity was also assessed in the same way. The rates of rash, hemorrhage, nausea, vomiting and taste disturbance increased as dose increased (P < 0.05), whereas the incidence of headache, abdominal pain, edema, fatigue, anemia, infection, muscle cramp and constipation was nearly identical and showed no significant difference. CONCLUSIONS: Imatinib at a standard dose produces a similar effect to that at a high dose. The severity of the toxicity is associated with the dose of imatinib. However, larger and randomized studies are needed to draw definitive conclusions.

Serum carbohydrate antigen (CA) 19-9 as a prognostic factor in cholangiocarcinoma: a meta-analysis.

This study was performed to determine the prognostic role of preoperative serum carbohydrate antigen (CA) 19-9 levels in the survival of patients with cholangiocarcinoma. Articles published up to June 1(st), 2010 that evaluated preoperative CA19-9 levels and the prognosis of cholangiocarcinoma were collected for meta-analysis. The required information for calculating individual relative risk (RR) was extracted from the studies, and a combined overall RR was estimated. Nine eligible studies were included. One study dealt with extra-hepatic cholangiocarcinoma, while the other eight studies analyzed intra-hepatic cholangiocarcinoma. The mean methodological quality score was 74.1%, ranging from 65.5% to 82.5%. The overall RR for the nine studies was 1.28 (95% confidence interval = 1.10-1.46), and the Z-score for overall effect was 13.83 (P<0.001). The association between serum CA19-9 level and lymph node involvement was also assessed. The combined RR was 1.471 (95% confidence interval = 0.411-5.264) and Z-score for overall effect was 0.59 (P = 0.553). CA19-9 levels were associated significantly with the prognosis of patients with cholangiocarcinoma. This meta-analysis shows that elevation of preoperative CA19-9 levels is correlated with a poor prognosis of patients with cholangiocarcinoma. However, larger scale and randomized studies are needed to draw a more substantive conclusion.