摘要
目的诱导构建人胃癌奥沙利铂(oxaliplatin,L-OHP)耐药细胞系SGC7901/L-OHP,探讨RNAi沉默基质细胞衍生因子-1(stromal cell-derived factor-1,SDF-1)表达对SGC7901/L-OHP增殖、凋亡、耐药的影响。方法L-OHP高浓度间歇冲击法诱导构建耐药细胞系SGC7901/L-OHP,观察其生物学行为变化,RT-PCR及Western blot法检测SDF-1 mRNA和蛋白表达情况;RNAi沉默SGC7901/L-OHP细胞SDF-1基因,从mRNA和蛋白水平验证沉默效果,MTT法及流式细胞术检测SGC7901/L-OHP细胞增殖、凋亡及细胞周期变化。结果成功建立耐药细胞系SGC7901/L-OHP,L-OHP对其增殖抑制率明显低于SGC7901细胞,其G 0/G 1期细胞增多、S期细胞减少,且SDF-1 mRNA和蛋白表达量明显高于SGC7901细胞(P均<0.05)。成功沉默SDF-1基因表达后,与空白及阴性对照组比较,SDF-1-RNAi组细胞增殖抑制率明显升高,对L-OHP药物敏感度明显升高,且其细胞凋亡也明显升高,G 0/G 1期细胞明显减少,S期细胞明显增多(P均<0.05)。结论本实验成功建立人胃癌L-OHP耐药细胞系SGC7901/L-OHP,并发现SDF-1参与胃癌细胞SGC7901对L-OHP获得性耐药过程;RNAi沉默SDF-1基因表达后抑制SGC7901/L-OHP细胞增殖、促进凋亡并部分逆转其对L-OHP的耐药性。
Objective To induce and establish the oxaliplatin resistant human gastric cancer cell line SGC7901/L-OHP,and investigate the effect of RNAi-silenced stromal cell-derived factor-1(SDF-1)gene on proliferation,apoptosis and durg resistance ofSGC7901/L-OHP.Methods Human gastric cancer SGC790l cells were intermittently exposed to L-OHP in manner high concentration impact in vitm to induce and establish L-OHP-resistant cell line SGC7901/L-OHP,then observe the changes of its biological behavior.RT-PCR and Western blot were used to detect the mRNA and protein expressions of SDF-1,respectively.The expression of SDF-1 gene in SGC7901/L-OHP was silenced by RNAi technology and the silencing efficiency was validated at both mRNA and protein level.MTT assay and flow cytometry were used to detect the proliferation,apoptosis and cell cycle changes of SGC7901/L-OHP cells.Results The L-OHP resistant cell line SGC7901/L-OHP was successfully established.The inhibition rate of L-OHP on its proliferation was significantly lower than that in SGC7901 cells,and its G 0/G 1 phase cells increased,S phase cells decreased,and the mRNA and protein expression of SDF-1 were significantly higher than that of SGC7901 cells(all P<0.05).The inhibition rate of cell proliferation in SDF-1-RNAi group was significantly higher than that in blank and negative control group,the drug sensitivity to L-OHP was increased significantly,and the apoptotic rate was also significantly increased.The G 0/G 1 phase cells were significantly reduced,meanwhile the S phase cells were significantly increased in SDF-1-RNAi group(all P<0.05).Conclusion In this study,human gastric cancer L-OHP resistant cell line SGC7901/L-OHP was successfully established,and SDF-1 was found to be involved in the acquired resistance of gastric cancer cell SGC7901 to L-OHP.RNAi-silenced SDF-1 gene can inhibit SGC7901/L-OHP cell proliferation,promote apoptosis and partly reverse its resistance to L-OHP.
作者
刘小慧
贺曼曼
张伟
李静
郭明海
冯运章
Liu Xiaohui;He Manman;Zhang Wei(General SurgeryⅡ,Handan Central Hospital,Hebei 056001,China)
出处
《医学研究杂志》
2019年第10期97-103,共7页
Journal of Medical Research
基金
河北省重点研发计划自筹项目(172777201)
