摘要
目的:探讨健脾补肾解毒方(JBJR)对大肠癌细胞转移及其对上皮-间质转化的作用。方法:1设健脾补肾解毒方醇提物0、30、60、90、120、150、180、210、240μg/ml浓度组,以CCK-8法检测各组对人大肠癌Lo Vo细胞增殖的抑制作用。2设空白细胞组、健脾补肾解毒方低剂量组(30μg/ml)、健脾补肾解毒方中剂量组(60μg/ml)、健脾补肾解毒方高剂量组(120μg/ml)、5-Fu组(15μg/ml)及桂枝汤醇提物组(120μg/ml),以Transwell法检测各组对大肠癌Lo Vo细胞侵袭转移的影响。3采用Real time PCR、Western blot、ELISA分别检测空白细胞组及健脾补肾解毒方低、中、高剂量组相关基因、特征蛋白和侵袭转移相关因子的表达情况。结果:1健脾补肾解毒方醇提物对大肠癌Lo Vo细胞有明显的增殖抑制作用,并呈时间和剂量依赖性,48 h的半数抑制浓度IC50为120μg/ml。2健脾补肾解毒方醇提物可不同程度地抑制Lo Vo细胞的转移能力,呈明显的剂量依赖性;其中高剂量组与5-Fu组的作用无统计学差异(P>0.05),而桂枝汤组无明显的抑制作用。3健脾补肾解毒方作用Lo Vo细胞48 h后,可剂量依赖性地上调E-cadherin mRNA、Snail及E-cadherin蛋白表达,下调Snail mRNA与Vimentin蛋白表达,实现抑制Lo Vo细胞的上皮-间质转化;可以抑制Lo Vo细胞分泌MMP-2、MMP-9。结论:健脾补肾解毒方醇提物可抑制大肠癌细胞转移,抑制上皮-间质转化和转移相关因子。
Objective： To discusses the impact and mechanism of Jianpi Bushen Jiedu Recipe（ JBJR） on the metastasis and Epithelial-to-Mesenchymal Transition（ EMT） of colorectal cancer cells. Methods： 1 Various concentrations of JBJR alcohol extract were set as 0,30,60,90,120,150,180,210 and 240 μg / ml,and then the inhibition on the proliferation of CRC Lo Vo cells was examined using CCK-8 method. 2The impact of different groups（ control group,JBJR groups with different concentrations at 30 μg / ml,60 μg / ml and 120 μg / ml,5-Fu group with concentration at 120 μg / ml,Guizhi decoction with concentration at 120 μg / ml） on the metastasis and invasion of CRC Lo Vo cells were accessed and compared using Transwell method. 3The expression of associated genes,characteristic proteins and related factors of invasion and metastasis to different groups（ blank group,JBJR groups with different concentrations at 30 μg / ml,60 μg / ml and 120 μg / ml） were examined using Real time PCR,Western blot and ELISA,respectively. Results： 1 JBJR alcohol extract can significantly inhibit the proliferation of Lo Vo cells depending on the dose and treatment duration of this recipe, The half maximal inhibitory concentration（ IC50） within 48 h is 120 μg/ml. 2 JBJR alcohol extract can significantly inhibit the migration ability of Lo Vo cells depending on the dose of this recipe; the effects between high dose group（ 120 μg / ml） and 5-Fu group are similar（ P 0. 05）,whereas the Guizhi decoction group showed low inhibition effect. 3 EMT can be dose-dependently inhibited through increasing the protein expression of E-cadherin mRNA,Snail and E-cadherin and the repressing the protein expression of Snail mRNA and Vimentin when the Lo Vo cells cultured with JBJR after 48 h. JBJR can inhibit the secretion of MMP-2 and MMP-9 in Lo Vo cells as well. Conclusion： JBJR alcohol extract was demonstrated to inhibit the migration of CRC cells and inhibit the associated factors of metastasis and EMT process.
出处
《上海中医药大学学报》
CAS
2016年第2期46-51,共6页
Academic Journal of Shanghai University of Traditional Chinese Medicine
基金
国家自然科学基金资助项目(81303102
81303103
81473478
81473628
81573749)
上海市卫生局面上项目(20114037)
上海市卫计委青年科研基金项目(20154Y0001)
关键词
健脾补肾解毒方
大肠癌
转移
上皮-间质转化
体外实验
＂Jianpi Bushen Jiedu Recipe＂
colorectal cancer
metastasis
Epithelial-to-Mesenchymal Transition
in vitro
