标题：YWHAE表达沉默对胃癌MGC803细胞增殖的影响

      作者：黄幼生，解娜，张艺馨，孙芳娇，罗志飞，薛逢贵    (海南医学院第一附属医院病理科，海南 海口 571101)

      卷次： 2017年28卷16期

      【摘要】 目的 探讨沉默 YWHAE表达对胃癌细胞增殖的影响。方法 实验分为三组，未转染组，构建YWHAE短发夹RNA (shRNA)表达载体及相应空载载体，慢病毒包装后分别感染胃癌MGC803细胞(干扰组，对照组)，应用Western blot、RT-PCR方法检测YWHAE沉默效率；MTT技术观察YWHAE表达沉默前后细胞增殖变化；流式细胞术、细胞克隆实验检测细胞周期、凋亡及克隆形成变化情况。结果 慢病毒包装YWHAE-shRNA后成功感染胃癌MGC803细胞；RT-PCR、Western blot检测结果显示，与对照组比较，干扰组MGC803细胞YWHAE基因表达明显降低，抑制效率为 72.7%。MTT结果显示，干扰组细胞增长速率明显低于对照组细胞，72 h增长倍数分别为1.56、2.43，差异有显著统计学意义(P<0.01)；细胞克隆实验结果显示，干扰组及对照组克隆形成率分别为 6.24%、31.00%，差异有显著统计学意义(P<0.01)。流式细胞术检测显示，YWHAE基因表达沉默后，MGC803细胞凋亡率增加，由未消减前的3.23%增加到9.83%，差异有显著统计学意义(P<0.01)；相比对照组细胞在G1期的比例(57.88%)，干扰组G1期细胞比例明显增加(72.42%)，差异有显著统计学意义(P<0.01)。结论 沉默YWHAE表达能抑制胃癌MGC803细胞增殖，可能与诱导细胞周期阻滞，促进细胞凋亡有关。
      【关键词】 YWHAE；胃癌；RNA干扰；细胞增殖；凋亡
      【中图分类号】 R735.2 【文献标识码】 A 【文章编号】 1003—6350（2017）16—2581—05

Silencing of YWHAE expression suppresses the proliferation ability of gastric cancer MGC803 cells.
HUANGYou-sheng, XIE Na, ZHANG Yi-xin, SUN Fang-jiao, LUO Zhi-fei, XUE Feng-gui. Department of Pathology, the FirstAffiliated Hospital of Hainan Medical University, Haikou 571101, Hainan, CHINA
【Abstract】 Objective To investigate the effect of YWHAE expression silencing on the proliferation of gastriccancer cells. Methods The expression vector of short hairpin RNA (shRNA) sequences for YWHAE (interferencegroup) and the control vector (control group) were constructed, which were used to infect the gastric cancer cellsMGC803 after they were produced and packaged by lentiviruses. The expression levels of YWHAE were assayed by West-ern blot and RT-PCR. Proliferation was evaluated by MTT (3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyl-tetrazoliumbromide) colorimetric assay. Colony formation analysis was used to determine growth properties of transduced cells.Cell cycle and apoptosis were determined by flow cytometry. Results Lentivirus-mediated shRNA-YWHAE was suc-cessfully transfected into gastric cancer cells. Compared with the control group, the expression of YWHAE was signifi-cantly decreased by 72.7% in the interference group by Western blot, RT-PCR analysis. The results of MTT showed thatthe proliferation ability of the interference group cells was significantly slower than that of the control group, and the cellgrowth times were 1.56 and 2.43 for 72 h, respectively (P<0.01). Cell cloning experiment results showed that the cloneformation rate of the interference group was 6.24%, which was significantly lower than 31.00% of the control group (P<0.01). Flow cytometry showed that reduced YWHAE expression in gastric cancer cells lead to the increasing of cell apop-tosis rate (from 3.23% to 9.83%), and the difference was statistically significant (P<0.01). Compared with the controlgroup (57.88%), the proportion of the interference group cells (72.42%) in G1 phase increased significantly (P<0.01).Conclusion The expression silencing of YWHAE inhibits the proliferation ability of gastric cancer MGC803 cells,which may be associated with cell apoptosis promotion and cell cycle G1 arrest.
      【Key words】 YWHAE; Gastric cancer; RNA interference (RNAi); Proliferation; Apoptosis·论 著·doi:10.3969/j.issn.1003-6350.2017.16.001基金项目: 国家自然科学基金(编号：81260321)