miR-135b-5p靶向CMTM3调节胃癌细胞的增殖、侵袭和迁移能力

目的:探究miR-135b-5p通过靶向CMTM3基因调节胃癌细胞的增殖、侵袭和迁移能力。方法:采用实时荧光定量PCR（RT-qPCR）检测胃癌组织和细胞系中CMTM3和miR-135b-5p的表达水平。利用慢性病毒转染技术将miR-135b-5p inhibitor转染至胃癌细胞，RT-qPCR检测转染效率；CCK-8比色法和Transwell实验检测转染后细胞的增殖、侵袭和迁移能力；Western blotting 检测转染后细胞中CMTM3蛋白表达水平。利用生物信息学网站预测miR-135b-5p潜在靶基因CMTM3，利用荧光素酶实验进行验证。结果:RT-qPCR检测结果表明，miR-135b-5p在胃癌组织和胃癌细胞株中呈现高表达，CMTM3在胃癌组织中呈现低表达（P<0.01）。RT-qPCR检测转染效率,结果显示，转染miR-135b-5p inhibitor敲低了miR-135b-5p的表达水平（P<0.001）；CCK-8和Transwell实验结果表明，敲低miR-135b-5p后抑制了胃癌细胞的增殖、侵袭和迁移能力（P<0.01），Western blotting实验结果表明，敲低miR-135b-5p促进了CMTM3蛋白的表达。生物信息学网站预测CMTM3为miR-135b-5p的潜在靶基因，荧光素酶实验证实了miR-135b-5p直接靶向胃癌细胞中CMTM3基因（P<0.01）。结论:miR-135b-5p作为重要的调节因子，反向调节抑癌靶基因CMTM3，从而促进胃癌细胞的增殖、侵袭和迁移能力。

miR-135b-5p targets CMTM3 to regulate the proliferation,invasion and migration of gastric cancer cells

Objective:To explore the ability of miR-135b-5p to regulate the proliferation,invasion and migration of gastric cancer cells by targeting the CMTM3 gene.Methods:The expression of CMTM3 and miR-135b-5p in gastric cancer tissues and cells was detected by RT-qPCR.miR-135b-5p inhibitor was transferred into gastric cancer cells by chronic virus transfection technique，and the transfection efficiency was detected by RT-qPCR.CCK-8 colorimetric assay and Transwell test were used to detect the proliferation，invasion and migration ability of gastric cancer cells transfected with miR-135b-5p inhibitor，respectively.The expression level of CMTM3 protein after transfection was detected by Western blotting.The potential target gene CMTM3 of miR-135b-5p was predicted by the biological information website，and the luciferase experiment was used to verify the results.Results:The results of RT-qPCR showed that miR-135b-5p was highly expressed in gastric cancer tissue and gastric cancer cells，and CMTM3 showed low expression in gastric cancer tissue(P<0.01).The results of RT-qPCR detection of transfection efficiency showed that the expression level of miR-135b-5p was decreased by transfection of miR-135b-5p inhibitor(P<0.001).The results of vitro transfection experiments showed that the proliferation，invasion and migration ability of gastric cancer cells were inhibited by knockdown of miR-135b-5p(P<0.01).The results of Western blotting assay showed that knockdown of miR-135b-5p promoted the expression of CMTM3 protein.The bioinformatics website predicted that CMTM3 was a potential target gene of miR-135b-5p，and the fluciferase test confirmed that miR-135b-5p was directly targeted at CMTM3 gene(P<0.01).Conclusion:As an important regulatory factor，miR-135b-5p inversely regulates the target gene CMTM3，which promotes the proliferation,invasion and migration of gastric cancer cells.