摘要
目的:探讨加味香砂六君子汤含药血清对胃癌细胞SGC7901增殖、侵袭的影响及其机制。方法:20只Wistar大鼠随机分为空白组,加味香砂六君子汤低剂量组、中剂量组、高剂量组[(1.55、3.1、6.2 g·(kg·d)-1]及5-FU组(0.5 g·L-1),每组5只。对照组灌服等体积生理盐水,按常规方法动物等体积灌胃,每天2次,连续5 d。末次灌胃后1 h固定动物,按无菌操作进行动物颈动脉采血,分离血清,56℃,经0.22μm滤膜抽滤除菌,制备低剂量组、中剂量组、高剂量组含药血清干预人胃癌SGC7901细胞。CCK8法观察不同浓度含药血清对细胞增殖影响;Transwell细胞侵袭实验检测含药血清作用SGC7901后细胞侵袭能力;梯度PCR检测Smo、Ptch1、Gli1的mRNA表达;蛋白质印迹法(Western blot)检测Smo、Ptch1、Gli1蛋白表达水平。结果:不同浓度含药血清对SGC7901增殖有明显抑制作用,随着浓度的增加,抑制率更加明显,且各实验组之间比较,差异具有统计学意义(P<0.05)。与对照组比较,各浓度含药血清作用后SGC7901细胞穿膜细胞数降低(P<0.05);与对照组比较,不同浓度的含药血清可明显降低Smo、Ptch1、Gli1的mRNA表达(P<0.05);各实验组Smo、Ptch1、Gli1蛋白表达也明显减少(P<0.05)。结论:加味香砂六君子汤含药血清对胃癌细胞增殖有抑制作用,抑制细胞侵袭,其机制可能与抑制Hh信号通路活化有关。
Objective:To investigate the effect of Jiawei Xiangsha Liujunzi Decoction serum on the proliferation and invasion of gastric cancer cell line SGC7901 and its mechanism.METHODS:Twenty Wistar rats were randomly divided into a blank group,and the low-dose group,the middle-dose group,and the high-dose [(1.55,3.1,6.2 g·(kg·d)-1] group and 5-FU(0.5 g·L-1) group,5 in each group.The control group was given the same volume of normal saline,and the rats were orally administered in the same volume as usual,twice a day for 5 days.The animals were fixed 1 hour after the last gavage.According to the aseptic operation,the blood was collected from the carotid artery of the animal,and the serum was separated and sterilized by filtration through a 0.22 μm filter at 56 ℃ to prepare a low-dose group,a middle-dose group and a high-dose group containing serum to interfere with human gastric cancer SGC7901 cells.With CCK8 method,the effect of serum containing different concentrations on cell proliferation was observed.Transwell cell invasion assay was used to detect the invasive ability of drug-containing serum after SGC7901.The mRNA expression of SMO,PTCH1 and GLI1 was detected with gradient PCR,SMO and PTCH1 and GLI1 protein expression level were detected with Western blot.Results:Different concentrations of drug-containing serum significantly inhibited the proliferation of SGC7901.With the increase of concentration,the inhibition rate was more obvious,and the difference between the experimental groups was statistically significant(P<0.05).Compared with the control group,the number of trans-membrane cells of SGC7901 cells decreased after each concentration of drug-containing serum(P<0.05).Compared with the control group,different concentrations of drug-containing serum could significantly reduce the expression of m RNA of SMO,PTCH1 and GLI1(P<0.05,the expression of SMO,PTCH1 and GLI1 protein in each experimental group was also significantly decreased(P<0.05).Conclusion:Jiawei Xiangsha Liujunzi Decoction-containing serum can inhibit the proliferation of gastric cancer cells and inhibit cell invasion.The mechanism may be related to inhibition of Hh pathway activation.
作者
杨君
邢向荣
高宏
殷东风
YANG Jun;XING Xiangrong;GAO Hong;YIN Dongfeng(Liaoning University of Traditional Chinese Medicine,Shenyang Liaoning China 110032;The Affiliated Hospital to Liaoning University of Traditional Chinese Medicine,Shenyang Liaoning China 110032)
出处
《中医学报》
CAS
2019年第10期2164-2169,共6页
Acta Chinese Medicine
基金
辽宁省中医药临床学(专)科能力建设资助项目(2013-Lnzyxzk-01)
关键词
加味香砂六君子汤
胃癌
细胞增殖
细胞侵袭
Hh信号通路
modified Xiangsha Liujunzi decoction
gastric cancer
cell proliferation
invasion
Hedgehog signaling pathway
