miR-601在胃癌中的表达及其对胃癌细胞凋亡、侵袭的影响

Expression of miR-601 in gastric cancer and its effect on the apoptosis and invasion of gastric cancer cells

目的探讨miR-601在胃癌中的表达及其对胃癌细胞凋亡、侵袭的影响。方法选取2018年8月至2019年8月台州市立医院收治的经病理学检查确诊为胃癌的30例患者的癌组织及相应癌旁正常组织标本;用转染试剂Lipofectamine 3000将miR-601抑制物(miR-601 inhibitor)和阴性对照序列(inhibitor NC)转染人胃癌细胞株AGS,并分为miR-601 inhibitor组和inhibitor NC组。采用qRT-PCR法检测miR-601在胃癌组织、癌旁正常组织、人胃癌细胞株AGS、HGC-27、MGC-803、正常胃细胞株GES-1以及miR-601 inhibitor组和inhibitor NC组中的相对表达量;采用MTT法检测两组细胞抑制率;采用流式细胞术检测两组细胞凋亡率;采用细胞划痕实验和Transwell小室实验检测两组细胞迁移率和细胞侵袭数;采用Western bolt法检测两组细胞基质金属蛋白酶(MMP)-2、MMP-9蛋白相对表达量。结果与癌旁正常组织比较,胃癌组织中miR-601相对表达量明显升高(P<0.01);与GES-1比较,其余3株胃癌细胞miR-601相对表达量均明显升高(均P<0.05);与inhibitor NC组比较,miR-601 inhibitor组miR-601相对表达量明显升高,转染miR-601 inhibitor后24、48、72 h时细胞抑制率均较低,细胞凋亡率明显增加,细胞迁移率明显降低,细胞侵袭数明显减少,MMP-2和MMP-9蛋白相对表达量均明显降低(均P<0.05)。结论miR-601在胃癌组织及细胞系中表达升高,下调其表达水平后可促进胃癌细胞凋亡。

Objective To investigate the expression of miR-601 in gastric cancer and its effect on the apoptosis and invasion of gastric cancer cells.Methods The cancer tissue and corresponding normal tissue specimens of 30 patients diagnosed with gastric cancer in Taizhou Municipal Hospital from August 2018 to August 2019 were collected.Human gastric adenocarcinoma AGS cells were transfected with miR-601 inhibitor and negative control sequence(NC group).The qRT-PCR method was used to detect the relative expression level of miR-601 in gastric cancer tissues,normal adjacent tissues,gastric cancer cell lines AGS,HGC-27,MGC-803,normal gastric cell lines GES-1.MTT method was used to detect the cell inhibition rate,flow cytometry was used to detect the apoptosis rate,the cell migration rate and cell invasion number were detect by the cell scratch assay and the Transwell chamber assay.Western bolt was used to detect the relative expression of matrix metalloproteinase 2 and 9(MMP-2,9).Results Compared with normal tissues,the relative expression of miR-601 in gastric cancer tissue was significantly increased(P<0.01),compared with GES-1 cells,the relative expression of miR-601 was significantly increased in gastric cancer AGS,HGC-27 and MGC-803 cells(P<0.05).Compared with the NC group,the relative expression of miR-601 in the miR-601 inhibitor group was significantly increased.The cell inhibition rate was reduced at 24,48,and 72 h after miR-601 inhibitor transfection,and the apoptosis rate was significantly increased.The migration rate and the number of cell invasion were reduced,and the relative expression of MMP-2 and MMP-9 proteins were significantly reduced(P<0.05).Conclusion The expression of miR-601 is up-regulated in tissues and cell lines of gastric cancer,and down-regulating miR-601 expression may inhibit proliferation and promote apoptosis of gastric cancer cells.