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miR-139在结肠癌、肝癌、外周神经鞘瘤、食管鳞状细胞癌、胶质瘤、胃癌中低表达,表明其在上述各种肿瘤中起到重要作用。本研究旨在验证miR-139在胃癌组织中的表达情况,就其在胃癌发生发展中的作用机制进行初步研究。采用实时荧光定量RT-PCR分析胃癌和癌旁组织中miR-139的表达水平;运用生物信息学手段,预测miR-139靶基因,并采用RT-PCR和Western blot分别检测潜在的靶基因在胃癌组织中的mRNA表达和蛋白水平表达变化;将miR-139 mimics瞬时转染胃癌细胞AGS细胞系,RT-PCR检测靶基因的表达水平。对瞬时转染miR-139 mimics及inhibitors的AGS细胞系,MTT法检测细胞增殖情况,流式细胞术检测细胞周期变化,Tran swell实验检测细胞侵袭能力。结果:(1)miR-139在胃癌中低表达。(2)生物学预测ROCK2是miR-139的靶基因,其在胃癌组织中mRNA和蛋白高表达。(3)miR-139 mimics转染后,抑制ROCK2 mRNA表达。(4)miR-139对胃癌的抑制作用:MTT结果表明,miR-139可抑制胃癌细胞增殖。Tran swell实验提示miR-139可抑制细胞的侵袭,miR-139可能通过下调靶基因的表达抑制胃癌细胞的增殖、侵袭能力。
miR-139 is low expressed in colon, liver, peripheral schwannoma, esophageal squamous cell carcinoma, glioma and gastric cancer, indicating that miR-139 plays an important role in the above-mentioned various tumors. The purpose of this study was to validate the expression of miR-139 in gastric cancer and to investigate its mechanism in the development of gastric cancer. The expression of miR-139 was detected by real-time fluorescence quantitative RT-PCR in gastric cancer tissues and paracancerous tissues. Bioinformatics methods were used to predict the expression of miR-139 and RT-PCR and Western blot were used to detect the potential target genes in gastric cancer The mRNA and protein expression of miR-139 mimics were transiently transfected into the AGS cell line of gastric cancer cells. The expression of target gene was detected by RT-PCR. The AGS cell lines transfected with miR-139 mimics and inhibitors were detected by MTT assay. Cell cycle was detected by flow cytometry. Tran swell assay was used to detect cell invasion. Results: (1) miR-139 was low expressed in gastric cancer. (2) Biological Prediction ROCK2 is a target gene of miR-139, which is highly expressed in gastric cancer tissues. (3) After transfected with miR-139 mimics, the expression of ROCK2 mRNA was inhibited. (4) The inhibitory effect of miR-139 on gastric cancer: MTT results show that miR-139 can inhibit the proliferation of gastric cancer cells. Tran swell experiments suggest that miR-139 can inhibit cell invasion, miR-139 may inhibit the proliferation of gastric cancer cells by reducing the expression of target genes, invasion.