叉头蛋白3对肺腺癌细胞增殖和化疗药物顺铂敏感性的影响

doi:10.13481/j.1671-587x.20190612

摘要/Abstract

摘要： 目的：观察叉头蛋白3（FOXP3）在人肺腺癌组织中的表达及其对肺腺癌细胞增殖和顺铂敏感性的影响，阐明肺腺癌细胞对顺铂耐药的相关机制。方法：收集肺腺癌患者术后石蜡标本50例和正常肺组织10例。免疫组织化学染色方法检测肺腺癌组织和正常肺组织中FOXP3和Ki-67的表达，分析肺腺癌组织和正常肺组织中FOXP3阳性表达率的差异，Pearson相关分析法分析FOXP3和Ki-67表达的相关性。选取对数生长期的人肺腺癌A549细胞进行siRNA转染，将A549细胞分为Mock组（转染脂质体）、Control-siRNA组（转染Control-siRNA）和FOXP3-siRNA组（转染FOXP3-siRNA）。RT-qPCR法检测各组A549细胞中FOXP3 mRNA表达水平，Western blotting法检测各组A549细胞中FOXP3蛋白表达水平，CCK-8法检测各组A549细胞增殖活性及不同浓度（0、0.63、1.25、2.50和5.00mg·L^-1）顺铂作用后各组A549细胞生长抑制率，RT-qPCR和Western blotting法检测顺铂作用后各组A549细胞中FOXP3 mRNA和蛋白表达水平。结果：FOXP3在肺腺癌和正常肺组织的阳性表达率分别为62.0%和13.3%，组间比较差异有统计学意义（P<0.05）；FOXP3和Ki-67在肺腺癌组织中表达呈正相关关系（r=0.370，P<0.01）；FOXP3-siRNA组A549细胞中FOXP3 mRNA和蛋白表达水平均明显低于Mock组和Control-siRNA组（P<0.01）；顺铂作用48和72 h后，FOXP3-siRNA组A549细胞增殖活性明显低于Mock组和Control-siRNA组（P<0.05）；1.25、2.50和5.00 mg·L^-1顺铂作用后，FOXP3-siRNA组A549细胞生长抑制率均高于Mock组和Control-siRNA组（P<0.05）；1.25和2.50 mg·L^-1顺铂组A549细胞中FOXP3 mRNA和蛋白表达水平均明显高于0 mg·L^-1顺铂组（P<0.05）。结论：沉默FOXP3可抑制肺腺癌细胞的增殖，增加肺腺癌细胞对顺铂的敏感性。

关键词: 肺腺癌, 叉头蛋白3, Ki-67, 顺铂, 化疗敏感性

Abstract: Objective: To observe the expression of forkhead box protein 3(FOXP3) in human lung adenocarcinoma tissue and the effects of FOXP3 on the cell proliferation and the chemosensitivity to cisplatin in the lung adenocarcinoma cells,and to elucidate the relevant mechanism of the resistance of lung adenocarcinoma cells to cisplatin. Methods: A total of 50 paraffin-embedded samples of cancer tissue of the patients with lung adenocarcinoma and 10 normal lung tissue samples were selected. The expressions of FOXP3 and Ki-67 in the lung adenocarcinoma and normal lung tissues were detected by immunohistochemistry,the difference in the positive expression rates of FOXP3 between lung adenocarcinoma and normal lung tissues was analyzed,and the correlation between the expressions of FOXP3 and Ki-67 was analyzed by Pearson correlation analysis.The human lung adenocarcinoma A549 cells at the logarithmic phase were transfected with siRNAs,and the A549 cells were divided into Mock group (transfected with liposome),Control-siRNA group (transfected with Control-siRNA) and FOXP3-siRNA group (transfected with FOXP3-siRNA).RT-qPCR and Western blotting methods were used to detect the expression levels of FOXP3 mRNA and protein in the A549 cells in various groups,and CCK-8 method was used to detect the proliferation activities of cells in various groups and the inhibitory rates of proliferation of the A549 cells in various groups after treated with 0,0.63,1.25,and 2.50 mg·L^-1 cisplatin;RT-qPCR and Western blotting methods were used to detect the expression levels of FOXP3 mRNA and protein in the A549 cells after treated with cisplatin. Results: The positive expression rates of FOXP3 in lung adenocarcinoma tissue and normal lung tissue were 62.0% and 13.3%,and the difference was statistically significant(P<0.05);the expression of FOXP3 in lung adenocarcinoma tissue was positively correlated with the expression of Ki-67 in lung adenocarcinoma tissue (r=0.370,P<0.01). The expression levels of FOXP3 mRNA and protein in the A549 cells in FOXP3-siRNA group were significantly decreased compared with Mock group and Control-siRNA group (P<0.01);the proliferation activities of A549 cells in FOXP3-siRNA group were significantly decreased compared with Mock group and Control-siRNA group after treated with cisplatin for 48 and 72 h (P<0.05);the inhibitory rates of proliferation of A549 cells in FOXP3-siRNA group were significantly higher than those in Mock group and Control-siRNA group after treatment with 1.25,2.50 and 5.00 mg·L^-1 cisplatin (P<0.05).The expression levels of FOXP3 mRNA and protein in the A549 cells in 1.25 and 2.50 mg·L^-1 cisplatin groups were both higher than those in 0 mg·L^-1 cisplatin group (P<0.05). Conclusion: Silencing FOXP3 can inhibit the levels proliferation and increase the chemosensitivity to cisplatin of the lung adenocarcinoma cells.

Key words: lung adenocarcinoma, forkhead box protein 3, Ki-67, cisplatin, chemosensitivity

中图分类号:

R734.2

房惠, 杨红宇, 马绪哲, 陈立松, 盖晓东, 历春. 叉头蛋白3对肺腺癌细胞增殖和化疗药物顺铂敏感性的影响[J]. 吉林大学学报(医学版), 2019, 45(06): 1261-1266.

FANG Hui, YANG Hongyu, MA Xuzhe, CHEN Lisong, GAI Xiaodong, LI Chun. Effects of FOXP3 on cell proliferation and chemosensitivity to cisplatin in lung adenocarcinoma cells[J]. Journal of Jilin University(Medicine Edition), 2019, 45(06): 1261-1266.

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