摘要
目的研究ANXA2-siRNA转染对胃癌耐药细胞药物敏感性及信号通路激活的影响。方法采用ANXA2-siRNA转染SGC-7901/DDP胃癌耐药细胞,通过RT-PCR和Western blot检测胃癌耐药细胞中耐药相关基因GST以及Bax的表达变化,应用MTT药敏实验检测胃癌耐药细胞对化疗药物敏感性的变化。采用Western blot检测ANXA2-siRNA转染SGC-7901/DDP胃癌耐药细胞后,MAPK及PI3K/Akt信号通路激活中P38、P38磷酸化水平、AKT以及AKT磷酸化水平的变化情况。结果 ANXA2-siRNA转染SGC7901/DDP细胞后,无论在RNA水平还是蛋白水平上,耐药相关基因GST的表达水平显著降低,而Bax的表达水平明显上调。与空白对照组和空白-siRNA组细胞相比,ANXA2-siRNA转染SGC7901/DDP细胞对化疗药物DDP和5-FU的IC50值显著降低,差异有统计学意义(P <0. 05)。与空白对照组和空白-siRNA组细胞相比,ANXA2-siRNA转染SGC7901/DDP细胞后,MAPK信号通路中,P38表达无明显变化,但P38磷酸化水平下降明显。在PI3K/Akt信号通路中,AKT表达无明显变化,但AKT磷酸化水平下降明显。结论 ANXA2-siRNA转染胃癌耐药细胞能够下调耐药相关基因的表达,降低胃癌细胞的耐药性,可能是通过影响MAPK及PI3K/Akt信号通路中P38和AKT磷酸化过程实现的。
Objective To explore the effect of ANXA2-siRNA transfection on drug sensitivity and signaling path-way activation in gastric cancer resistant cells.Methods ANXA2-siRNA was used to transfect SGC-7901/DDP gastric cancer drug-resistant cells.RT-PCR and Western blot were used to detect the expression of GST and Bax in drug-resistant gastric cancer cells.The chemotherapeutic drug sensitivity was detected in gastric cancer cells by MTT susceptibility test.Western blot was used to detect the changes of P38,P38 phosphorylation,AKT and AKT phosphorylation level after ANXA2-siRNA transfection in SGC-7901/DDP gastric cancer drug-resistant cells.Results After transfection of SGC7901/DDP cells with ANXA2-siRNA,the expression level of GST was significantly decreased at both RNA level and protein level,while the expression level of Bax was upregulated.Compared with blank control group and blank-siRNA group,ANXA2-siRNA transfected SGC7901/DDP cells significantly decreased the IC 50 value of chemotherapeutic drugs DDP and 5-FU,and the difference was statistically significant(P<0.05).Compared with blank control group and blank-siRNA group,ANXA2-siRNA transfected SGC7901/DDP cells did not change significantly in MAPK signaling pathway,but P38 phosphorylation level decreased significantly.Conculsion ANXA2-siRNA transfected gastric cancer drug-resistant cells can down regulate the expression of drug resistance related genes,and reduce the drug resistance of gastric cancer cells.It may be achieved by influencing the phosphorylation of P38 and AKT in MAPK and PI3K/Akt signaling pathways.
作者
余南荣
曾祥
徐厚巍
蓝俊松
Yu Nanrong;Zeng Xiang;Xu Houwei(Dept of Gastrointestinal Neoplasms,Cancer Center of Guangzhou Medical University,Guangzhou 510095)
出处
《安徽医科大学学报》
CAS
北大核心
2018年第11期1711-1715,共5页
Acta Universitatis Medicinalis Anhui
基金
广东省自筹经费类科技计划项目(编号:2017ZC0265)
关键词
ANXA2
转染
胃癌
耐药细胞
药物敏感性
信号通路
ANXA2
transfection
gastric cancer
drug-resistant cells
drug sensitivity
signal pathway
