粉防己碱通过FEZF1-AS1影响结直肠癌细胞化疗耐药的作用机制

Mechanism of tetrandrine affecting chemoresistance of colorectal cancer cells via FEZF1-AS1

目的探讨粉防己碱(tetrandrine,Tet)通过长链非编码RNA(LncRNA)FEZF1-AS1对结直肠癌细胞化疗耐药的作用机制。方法用浓度梯度实验筛选无毒剂量的Tet,建立人结直肠癌耐奥沙利铂(L-OHP)细胞株,设置空白组、Tet组、FEZF1-AS1组和FEZF1-AS1+Tet组。用MTT法检测各组细胞活力;用流式细胞术检测各组细胞凋亡率;用qRT-PCR检测各组细胞中化疗耐药相关基因ABCC1、MDR1、P-gp mRNA的表达水平;用Western blot检测各组细胞中ABCC1、MDR1、P-gp的表达水平。结果FEZF1-AS1组、空白组、FEZF1-AS1+Tet组和Tet组,HCT116/L-OHP细胞A_(490)值逐渐降低,凋亡率逐渐升高(P<0.05),ABCC1、MDR1、P-gp mRNA和蛋白的表达水平逐渐降低(P<0.05)。结论Tet可能通过下调LncRNA FEZF1-AS1降低结直肠癌细胞对化疗药物的耐药性。

Objective To investigate the mechanism of tetrandrine(Tet)on chemoresistance in colorectal cancer cells through long non-coding RNA(LncRNA)FEZF1-AS1.Methods The concentration gradient experiment was used to screen the Tet nontoxic dose.Oxaliplatin resistant human colorectal cancer cell lines were established and grouped into blank,Tet,FEZF1-AS1 and FEZF1-AS1+Tet.MTT assay was used to detect cell viability.The apoptosis rate was detected by flow cytometry.The mRNA and protein expression levels of ABCC1,MDR1 and P-gp were detected by qPCR and Western blot.Results In FEZF1-AS1,blank,FEZF1-AS1+Tet,and Tet groups the A_(490) value of HCT116/L-OHP cells gradually decreased,the apoptosis rate gradually increased(P<0.05),and the expression levels of ABCC1,MDR1,P-gp mRNA and protein gradually decreased(P<0.05).Conclusion Tet may reduce the resistance of colorectal cancer cells to chemotherapeutic drugs by downregulating LncRNA FEZF1-AS1.