摘要目的 观察微小RNA(miRNA,miR)-203通过SNAI2对胃癌细胞SGC7901侵袭和凋亡的影响.方法 (1)脂质体转染将miR-203 mimics转入胃癌细胞SGC7901,实时定量聚合酶链反应(Real-time PCR)检测转染效果,Transwell实验和流式细胞术检测细胞侵袭和凋亡.(2)Real-timePCR和Western blot检测转染miR-203 mimics后在胃癌细胞中SNAI2的表达水平.(3)小干扰RNA(siRNA)干扰胃癌细胞SGC7901中SNAI2的表达水平,Western blot验证敲减效果,并检测敲减SNAI2后细胞侵袭和凋亡.(4)转染miR-203 mimics后,脂质体转染SNAI2表达质粒,检测细胞侵袭和凋亡情况.结果 (1)胃癌细胞SGC7901转染miR-203 mimics后,miR-203的表达水平为6.34±0.73,对照组的表达水平为1.26±0.21,两组比较差异有统计学意义(t=1.528,P=0.024).(2)转染miR-203 mimics组穿透滤膜的细胞数明显少于对照组穿膜细胞数,两组比较差异有统计学意义(=1.781,P=0.016).(3)流式细胞术结果显示,对照组细胞凋亡率为(7.27±1.37)％,转染miR-203 mimics组胃癌细胞SGC7901的凋亡明显增加,为(38.62±6.27)％,两组比较差异有统计学意义(x2 =7.373,P =0.009).(4)转染SNAI2后,miR-203对胃癌细胞SGC7901的促凋亡作用被反转,转染SNAI2组的细胞凋亡率降低为(14.03±3.60)％,和miR-203 mimics组比较,差异有统计学意义(x2 =4.162,P =0.036).(5)与对照组比较,siRNA抑制SNAI2后,SGC7901细胞的凋亡明显增加,为(47.92±7.14)％,两组比较差异有统计学意义(x2=10.373,P=0.004).结论 miR-203能够通过靶向调控SNAI2而降低胃癌细胞SGC7901的侵袭能力并促进其凋亡.

abstractsObjective To investigate the effect of microRNA (miRNA,miR)-375 on cell invasion and apoptosis in the human gastric cancer cell line SGC7901.Methods The expression levels of miR-203 in the SGC7901 cells were determined by real-time quantitative polymerase chain reaction (Real-time PCR) after transfection of the miR-203 mimic.Transwell cell invasion and flow cytometry were used to detect the invasion and apoptosis of SGC7901 cells.then the expression levels of snail homolog 2 (SNAI2) in gastric cancer cell lines were assayed after transfection of the miR-203 mimic by Real-time PCR and Western blotting.the expression of SNAI2 was determined by Western blotting after knockdown by small interfering RNA (siRNA),and the invasion and apoptosis of SGC7901 cells after knockdown of SNAI2 was detected.finally,SNAI2 after transfection of miR-203 mimic was expressed,and the invasion and apoptosis of SGC7901 cells was detect.Results (1) After gastric cancer cells SGC7901 transfection miR-203 mimics,expression level of miR-203 was 6.34 ± 0.73,the miRNA con group the expression level of 1.26 ±0.21,comparing the two groups,the difference was statistically significant (t =1.528,P =1.528).(2) Transfection miR-203 mimics group through the membrane of cells was significantly less than transfection of miRNA con group through a membrane on the number of cells,comparing the two groups,difference was statistically significant (t =1.781,P =1.781).(3) Flow cytometry results showed that the miRNA con group the apoptosis rate was (7.27 ± 1.37) ％,transfection miR-203 mimics group of SGC7901 gastric cancer cells apoptosis increased significantly,as (38.62 ± 6.27) ％,comparing the two groups,difference was statistically significant (x2 =7.373,P =7.373).(4) After transfection SNAI2,miR-203 effect on promoting apoptosis of gastric cancer cells SGC7901 was reversed,transfection SNAI2 reduce the apoptosis rate in group is (14.03 ± 3.60) ％,and miR-203 mimics group comparison,statistically significant differences (x2 =4.162,P =4.162).(5) Compared with the control group,after siRNA SNAI2 inhibition,the apoptosis of SGC7901 cells increased significantly,for the (47.92 ±7.14)％,comparing the two groups,difference was statistically significant (x2 =10.373,P =10.373).Conclusion MiR-203 can inhibit invasion and promote apoptosis of SGC7901 gastric cancer cell by targeting SNAI2.