The Effect Of MiR-106b-5p On The Proliferation And Apoptosis Of Gastric Cancer Cells

Background:Gastric cancer is the fifth most common tumor in the world and the second leading cause of cancer deaths.Each year,new cases of gastric cancer rank fourth in the incidence of all tumors and women in the fifth.Of these new cases,more than 70% occurred in developing countries,and more than half occurred in East Asia(mainly in China).The epidemiology of gastric cancer in China is particularly serious.With the deepening of human understanding of cancer and the ever-increasing diagnostic techniques and treatment levels,the discovery rate of early-stage gastric cancer gradually increases and a better prognosis can be obtained.However,the long-term survival time of advanced gastric cancer is still not optimistic,and many factors such as genes Background,Helicobacter pylori infection,smoking can affect the occurrence and development of gastric cancer.Therefore,the study of advanced gastric cancer still needs to be deepened in order to obtain better clinical results.As the study of non-coding small RNA(miRNA)continues,its role in the development of tumors has also been revealed.In general,miRNAs can exert their effects on tumor cells through direct regulation of downstream target genes.At present,miR-106b-5p has been reported in liver cancer,renal clear cell carcinoma,non-small cell lung cancer,and glioma,which may regulate tumor cell proliferation,invasion and metastasis.However,whether miR-106b-5p is differentially expressed and whether it plays a role in gastric cancer cells is still at an unknown stage.This topic will focus on the biological effects and mechanisms of miR-106b-5p on tumors.Purpose:Reveal the expression level of miR-106b-5p in gastric cancer tissues and paired paraneoplastic tissues.Investigating the effect of miR-106b-5p on the proliferation and death of gastric cancer cells and its preliminary potential mechanism.Methods:1.The expression level of miR-106b-5p in 72 pairs of gastric cancer tissues and their paired adjacent tissues was analyzed by real-time quantitative PCR,and the correlation analysis was performed by combining the corresponding clinical data.The expression of miR-106b-5p in gastric cancer cell lines and normal gastric epithelial cell lines was determined by real-time quantitative PCR.Real-time Quantitative PCR Assay and Western Blot determination of expression Levels of mRNA and protein in target Genes.2.A stable lentivirus with specific miR-106b-5p overexpression and interference and a corresponding negative control were designed and transfected into gastric cancer cell lines MGC-803 and BGC-823.The CCK8 assay,plate colony formation assay,and EdU were used to determine the changes in cell proliferation and flow cytometry was used to determine the apoptotic rate and period distribution of the corresponding cells.3.Through a public database and PCR analysis,the target gene was predicted to be TXNIP.Using luciferase reporter assays,we tested whether our predicted target gene interacts with miR-106b-5p.Construct stable stably transduced and overexpressed TXNIP and the corresponding negative control and transfected BGC-823 and MGC-803 for rescue experiment.4.Subcutaneous injection of gastric cancer cells into the underarms of both sides of the nude mice to form a tumor,finally measuring the size of the tumor,so as to observe the proliferation of cells,implanted tumors in vivo removal,immunohistochemistry analysis of Ki-67 visually reflect the ability of tumor proliferation,immunohistochemistry Analysis of TXNIP expression in implanted tumors.Results: qRT-PCR showed that the expression of miR-106b-5p in gastric cancer tissues was significantly higher than that in adjacent tissues(P<0.01).Clinical data analysis showed that the expression of miR-106b-5p was related to the tumor size of gastric cancer.Positive correlation;CCK-8 assay,plate colony formation assay,and EdU assay found proliferation of MGC-803 cell lines after constructing an interference constructs for BGC-823 cell line and miR-106b-5p overexpressing for MGC-803 cell line.respectively.Enhanced,while the proliferation of BGC-823 cell line was weakened;the proportion of death of MGC-803 cell line was decreased by flow cytometry,and the cycle progress of BGC-803 cell line was accelerated.As a downstream target of miR-106b-5p,TXNIP can influence the biological function of gastric cancer cells.When transfected with TXNIP over-expression and interference with lentivirus,miR-106b-5p exerted a response to the function of gastric cancer cells.The experimental mouse xenograft model found that the miR-106b-5p interference group reduced the tumor volume of the mice and slowed the growth curve;Ki-67 staining showed a decrease in the proportion of nuclear staining,Western blot detection found that the miR-106b-5p interference group cells The expression of TXNIP was increased,while the overexpression group was the opposite;after transfection of TXNIP overexpression and interference with lentivirus,the above phenomenon achieved a recovery.Conclusions:The expression of MiR-106b-5p is positively correlated with tumor size and is highly expressed in gastric cancer tissues and cells.The highly expressed miR-106b-5p can increase the proliferation of cells through the regulation of the target gene TXNIP,thereby promoting the proliferation of gastric cancer cells.These results are consistent with the above-mentioned published miR-106b-5p expression levels and biological functions in various tumors with high reliability.In this study,the effect of miR-106b-5p on proliferation of gastric cancer cells has important clinical significance.