RNA干扰技术探讨参芪扶正注射液对MHCC97L细胞侵袭力的影响  

Effect of Shenqi Fuzheng injection on invasion of MHCC97L cells based on RNA interference technology

作　　者：陈晓珩[1] 何蓓 李会龙[1] 户蕊 李璐[1] 王鑫[1] 汪唐顺[1] 李乃卿[1] 丁治国[1] 

机构地区：[1]北京中医药大学东直门医院外一科,北京100700 [2]北京中医药大学第三附属医院医务处,北京100029

出　　处：《中国现代普通外科进展》2017年第7期505-510,共6页Chinese Journal of Current Advances in General Surgery

基　　金：国家自然科学基金(81673972);北京中医药大学自主选题项目(2016-JYB-JSMS-030)

摘　　要：目的:通过检测参芪扶正注射液对肝癌MHCC97L细胞侵袭力及其VEGF基因表达的影响,明确其降低肝癌瘤株侵袭力的作用,并通过RNA干扰技术加以验证。方法:在MHCC97L体外培养系中加入参芪扶正注射液(0.5 g/L)作用24 h后,Transwell检测参芪扶正注射液对MHCC97L细胞侵袭性的影响;同时使用Real-time PCR和Western-blot分别检测药物组、阴性对照组和空白对照组中VEGF基因的转录与表达情况,确定参芪扶正注射液对VEGF基因转录与表达的影响;构建VEGF干扰慢病毒载体并实现其在人肝癌细胞MHCC97L中表达,并通过RT-PCR、Western-Blot进行验证,确定其干扰VEGF基因表达的有效性;Transw ell检测参芪扶正注射液对VEGF"沉默"瘤株侵袭力的影响并与正常瘤株进行比较。结果:参芪扶正注射液能够降低MHCC97L细胞中VEGF基因的转录与表达(P<0.05),并能够降低肿瘤侵袭力(P<0.05);VEGF基因"沉默"瘤株构建成功;VEGF基因"沉默"后,参芪扶正注射液对MHCC97L瘤株侵袭力的影响不显著(P>0.05)。结论:参芪扶正注射液可以通过抑制MHCC97L细胞VEGF基因的表达以降低其侵袭力。Objective： To investigate the inhibitory effect of Shenqi Fuzheng injection on the expression of VEGF gene in human hepatocellular carcinoma cell line MHCC97L, and to elucidate the effect of Shenqi Fuzheng injection on tumor invasiveness, and to validate it through siRNA technolo- gy. Methods： After 24 hours of addition of Shenqi Fuzheng Injection （MHC） in the culture system of MHCC97L, Real-time PCR and Western-blot were used to detect the expression of MMP-9 and MMP-9 in the control group （Shenqi Fuzheng）, negative control group （NaCI） And the expression of VEGF and RNA in the blank control group, and to determine the effect of Shenqi Fuzheng injection on the expression of VEGF and RNA. The effect of Shenqi Fuzheng Injection on the invasiveness of MHCC97L cells was detected by Transwell assay. The lentiviral vectors of MHCC97L cells were transfected into 293T cells and transfected into MHCC97L cells. The transfected cells were transfected into 293T cells by Lipofectamine 2000, and transfected into MHCC97L cells. The Effect of Shenqi Fuzheng Injection on the Invasion of VEGFi Tumor by Transwell. Results： Shenqi Fuzheng injection could significantly decrease the expression of VEGF mRNA and protein in normal MHCC97L cells （P〈0.05）, and decrease the invasiveness of tumor cells （P〈0.05）. VEGF RNA interference was successful. After intervention, Shenqi Fuzheng injection could not decrease the invasiveness of MHCC97L tumor （P〉0.05）. Conclusion： Shenqi Fuzheng injection can reduce the invasion of MHCC97L cells by inhibiting the expression of VEGF gene.

关 键 词：肝肿瘤 参芪扶正注射液 VEGF 侵袭力 

分 类 号：R735.7[医药卫生—肿瘤]