六神丸逆转人结肠癌耐阿霉素细胞株Lovo/Dox机制的初步研究  

Preliminary mechanism research of Liushen Pill on reversing doxorubicin resistant human colon cancer cell line Lovo/Dox

作　　者：邹瑜[1] 袁玉霞[1] 贺雪[1] 唐雪瑶 张祎稀 邱艳艳[1] 袁泽婷 徐可[1] 殷佩浩[1] 

机构地区：[1]上海中医药大学附属普陀医院普外科,上海200062

出　　处：《上海中医药杂志》2017年第4期90-94,共5页Shanghai Journal of Traditional Chinese Medicine

基　　金：上海中医药大学"研究生创新"项目(2015YCX15);上海市普陀区卫计委"315"工程人才培养计划学科带头人后备人才资助项目(14Q-RC-08)

摘　　要：目的初步探讨六神丸逆转人结肠癌耐阿霉素细胞株Lovo/Dox的机制。方法设置对照组、阿霉素组(Dox)、六神丸组和阿霉素与六神丸联合用药组。用Cell Counting Kit-8(CCK-8)法检测六神丸对人结肠癌耐药细胞株(Lovo/Dox、HCT116/L-OHP、Caco2/Dox)的影响;蛋白质免疫印迹法(Western Blotting)检测耐药蛋白P-gp、BCRP以及相关凋亡蛋白caspase-3、caspase-9、Cytochrome C和抑癌蛋白p53在人结肠癌Lovo/Dox细胞中的表达;免疫荧光实验检测P-gp的荧光表达。结果六神丸作用于人结肠癌3种耐药细胞24 h后,CCK-8结果显示其可抑制3种人结肠癌细胞株的活性,其中抑制人结肠癌耐阿霉素细胞株Lovo/Dox的效果最明显。六神丸联合阿霉素可上调caspase-3、caspase-9、p53蛋白表达水平以及减弱P-gp蛋白及荧光表达。结论六神丸可逆转人结肠癌耐阿霉素细胞株Lovo/Dox的P-gp表达,可能是通过激活相关凋亡途径和抑癌蛋白而引起该细胞株凋亡。Objective To preliminarily discuss the mechanism of Liushen Pill on reversing doxorubicin resistant human culon cancer cell line Lovo/Dox. Methods The control group ,doxoruhiein group, Liushen Pill group and combination group were divided, The effects of Liushen Pill on drug-resistant human colon cancer cell lines （ Lovo/Dox, HCT116/L-OHP, Caco2/Dox） were detected hy Cell Counting Kit-8 （ CCK- 8 ）. The expressions of P-gp, BCRP and related apoptosis proteins caspase-3, caspase-9, Cytochrome C and tumor suppressor protein p53 in human colon cancer cell liue Lvo/Dox were determined by Western blot. The fluorescence expression of P-gp was detected by immunonuorescence assay. Results After the intervention of Liushen Pill or 24 hours, the CCK-8 showed that Liushen Pill could inhibit the activities of the three drug-resistant human colon cancer cell lines, among the inhibiting effects on doxorubicin resistaut human colon cancer cell line Lovo/Dox were most significant. The treatment of Liushen Pill combined with doxorubicin could up-regulate the protein expression of caspase-3, caspase-9 and p53 and decrease the protein and fluorescence expression of P-gp. Conclusion Liushen Pill can reverse the expression of P-gp in doxorubicin resistant human colon cancer cell line Lovo/Dox, which may be related to the activation of the related apoptosis pathway and tumor suppressor protein.

关 键 词：人结肠癌细胞株 六神丸 凋亡机制 蛋白表达 阿霉素 耐药逆转 

分 类 号：R285[医药卫生—中药学]