黄芪建中汤含药血清联合顺铂对人肺癌A549细胞增殖及E-cadherin、Snail1表达的影响

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摘要目的研究黄芪建中汤含药血清联合顺铂对人肺癌A549 细胞的作用及其对E-cadherin、Snail1 表达的影响。方法黄芪建中汤含药血清联合不同浓度顺铂（0、2、4、8、16、32、64 μg/ml）培养A549 细胞株48 h，采用四唑盐比色（MTT）法检测细胞株的增殖抑制率，确定联合顺铂的最佳用药浓度。将A549 细胞株随机分为4 组，即空白组、中药组、顺铂组、联合组，依次用完全培养基、黄芪建中汤含药血清、顺铂、黄芪建中汤含药血清联合顺铂干预。各组细胞株培养48 h 后，进一步采用MTT 法检测各组细胞株的增殖抑制率。再通过免疫细胞化学法（IHC）、免疫蛋白印迹（WB）法检测各组细胞株E-cadherin、Snail1 蛋白的表达。最后通过实时荧光定量PCR（RT-PCR）法检测各组细胞株E-cadherin、Snail1 基因的表达。结果 MTT 法研究显示，联合用药时顺铂的最佳浓度为8 μg/ml；联合组A549 细胞株增殖抑制率高于顺铂组、中药组、空白组，差异有统计学意义（P＜0.05）。IHC 和WB 法研究显示，联合组A549 细胞株中的E-cadherin 蛋白表达水平高于顺铂组、中药组、空白组，差异有统计学意义（P＜0.05）；联合组A549 细胞株中的Snail1 蛋白表达水平低于顺铂组、中药组、空白组，差异有统计学意义（P＜0.05）。RT-PCR 法研究显示，联合组A549 细胞株中的E-cadherin 基因表达水平高于顺铂组、中药组、空白组，差异有统计学意义（P＜0.05）；联合组A549 细胞株中的Snail1 基因表达水平低于顺铂组、中药组、空白组，差异有统计学意义（P＜0.05）。结论黄芪建中汤含药血清联合顺铂能够抑制A549 细胞株的增殖，增强人肺癌A549 细胞株中E-cadherin 蛋白、基因的表达，并降低人肺癌A549 细胞株中Snail1 蛋白、基因的表达。

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	王晓芳
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关键词 ：肺癌, A549 细胞, 黄芪建中汤, 顺铂, E-cadherin, Snail1

Abstract：Objective To study the effect of serum containing Huangqi Jianzhong Decoction combined with Cisplatin on A549 cell proliferation and expression of E-cadherin and Snail1 in human lung cancer.Methods A549 cell lines were cultured for 48 h with the serum containing Huangqi Jianzhong Decoction combined with different concentrations of Cisplatin(0,2,4,8,16,32,64 μg/ml).Magnetic Tape Terminal(MTT)was used to detect the proliferation inhibition rate of A549 cell lines and determine the optimal concentration of Cisplatin.A549 cell lines were randomly divided into four groups：blank group,traditional Chinese medicine group,Cisplatin group and combination group,in which complete medium,serum containing Huangqi Jianzhong Decoction,Cisplatin,serum containing Huangqi Jianzhong Decoction and Cisplatin were successively used for intervention.After A549 cell lines were cultured for 48 h,the proliferation inhibition rate of A549 cell lines in each group was further detected by MTT.The protein expression levels of Ecadherin and Snail1 in each group were determined by immunohistochemistry(IHC)and Western Blot(WB).The gene expression levels of E-cadherin and Snail1 were determined by realtime-PCR(RT-PCR).Results The research of MTT method showed that the optimal concentration of Cisplatin in combination was 8 μg/ml.The inhibition rate of A549 cell lines in the combination group was higher than that in the Cisplatin group,the traditional Chinese medicine group and the blank group,the differences were statistically significant(P＜0.05).The research of IHC and WB method showed that the expression level of E-cadherin protein of A549 cell lines in the combination group was higher than that in the Cisplatin group,the traditional Chinese medicine group and the blank group,the differences were statistically significant(P＜0.05).And the expression level of the Snail1 protein of A549 cell lines in the combination group was lower than that in the Cisplatin group,the traditional Chinese medicine group and the blank group,with statistically significant differences(P＜0.05).The research of RT-PCR method showed that the gene expression level of E-cadherin of the A549 cell lines in the combination group was higher than that in the Cisplatin group,the traditional Chinese medicine group and the blank group,with statistically significant differences(P＜0.05).The gene expression level of the Snail1 in the A549 cell lines of the combination group was lower than that of the Cisplatin group,the traditional Chinese medicine group and the blank group,the differences were statistically significant(P＜0.05).Conclusion Serum containing Huangqi Jianzhong Decoction combined with Cisplatin can inhibit the proliferation of A549 cell lines,enhance the expression of E-cadherin protein and gene in A549 cells line of human lung cancer,and reduce the expression of Snail1 protein and gene in A549 cells line of human lung cancer.

Key words： Lung cancer A549 cells Huangqi Jianzhong Decoction Cisplatin E-cadherin Snail1

基金资助:国家自然科学基金项目（81473569）

通讯作者: 刘春英（1962-），女，山东文登人，博士，教授，博士研究生导师，研究方向：中药抗肿瘤作用及其机制研究

作者简介: 王晓芳（1991-），女，山西吕梁人，辽宁中医药大学2017 级在读硕士研究生，研究方向：中药抗肿瘤作用及其机制研究

引用本文:

王晓芳；于丹；刘春英. 黄芪建中汤含药血清联合顺铂对人肺癌A549细胞增殖及E-cadherin、Snail1表达的影响[J]. 中国当代医药, 2020, 27(10): 4-8转12.
WANG Xiao-fang； YU Dan； LIU Chun-ying. Effect of serum containing Huangqi Jianzhong Decoction combined with Cisplatin on A549 cell proliferation and expression of E-cadherin and Snail1 in human lung cancer. 中国当代医药, 2020, 27(10): 4-8转12.

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http://dangdaiyiyao.com/CN/ 或 http://dangdaiyiyao.com/CN/Y2020/V27/I10/4

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