SHI Yan-ting, JING Huan, WANG Ying, et al. Rat Serum Containing of Buzhong Yiqi Decoction Reverse Chemotherapeutic Resistance in A549/DDP Cells by Down-regulating Expression Levels of mTOR[J]. Chinese journal of experimental traditional medical formulae, 2013, 19(9): 215-219.
SHI Yan-ting, JING Huan, WANG Ying, et al. Rat Serum Containing of Buzhong Yiqi Decoction Reverse Chemotherapeutic Resistance in A549/DDP Cells by Down-regulating Expression Levels of mTOR[J]. Chinese journal of experimental traditional medical formulae, 2013, 19(9): 215-219.DOI:
补中益气汤含药血清逆转A549/DDP的顺铂耐药及对mTOR表达的影响
摘要
目的:研究补中益气汤含药血清对肺腺癌顺铂耐药细胞株(A549/DDP)顺铂耐药的逆转作用及对哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin
mTOR)表达的影响。方法:动物随机分组
制备补中益气汤高、中、低剂量血清(给药剂量分别为11.34
5.67
2.83 g.kg-1.d-1)
将A549
A549/DDP细胞体外分为顺铂(DDP)组
空白血清+DDP组
含药血清高剂量+DDP组
含药血清中剂量+DDP组
含药血清低剂量+DDP组
细胞对照组
经过15%各组血清联合DDP不同浓度(10
20
40
80
100
200μmol.L-1)作用48 h
采用MTT法检测补中益气汤不同浓度含药血清对A549/DDP顺铂耐药的逆转作用;采用免疫荧光
免疫细胞化学
Western-blot方法检测mTOR的表达。结果:与DDP组比较
补中益气汤不同浓度含药血清作用于A549/DDP后IC50和耐药倍数均有显著降低(P<0.05)
以补中益气汤中剂量组(等临床剂量组)作用最为显著(P<0.01)
A549/DDP中mTOR的染色吸光度及荧光强度均明显减弱(P<0.01)
蛋白表达水平明显降低(P<0.01)。结论:补中益气汤能增强A549/DDP对顺铂的敏感性
并能通过降低细胞生存通路中mTOR的表达而有效逆转肺腺癌细胞的顺铂耐药。
Abstract
Objective:To identify the effect of rat serum containing different concentrations of Buzhong Yiqi decoction(BZYQT) on reverse the chemotherapeutic resistance in A549/DDP cells and on the expression of mTOR.Method: Animals were randomized to prepared the rat serum containing varied does levels of BZYQT(does for 11.34
5.67
2.83 g · kg-1 · d-1)
the A549
A549/DDP were equally divided into six groups in random: the DDP group
BZYQT low does+DDP group
BZYQT middle does+DDP group
BZYQT high does+DDP group
NS+DDP group and the cell control group.Followed by 15%serum of every group and the different concentrations of DDP(10
20
40
80
100
200 μmol · L-1) for 48 h
assayed by MTT to detect the rat serum containing of BZYQT reverse the chemotherapeutic resistance in A549/DDP.The expression of mTOR in cells were evaluated by indirect immunofluorescence
immunocytochemistry technique and Western-blot.Result: The median inhibitory concentration IC50 and resistant multiple of A549/DDP is significant reduction by the serum containing varied does levels of BZYQT(P<0.05)
especially the effect of the middle dose group is the most significant(P<0.01)
the optical density and fluorescence intensity of mTOR were significantly reduced(P<0.01) and the protein expression levels were significantly lower in the cells(P<0.01).Conclusion: BZYQT appeared to increase the chemotherapeutic sensitivity of A549/DDP cells
showing regression of multiple chemotherapeutic resistance in pulmonary adenocarcinoma by down-regulating expression levels of mTOR.