紫杉醇通过上调miR-7抑制非小细胞肺癌细胞的抗增殖作用并促进其凋亡的应用研究
|
摘要
目的研究紫杉醇通过上调miR-7抑制非小细胞肺癌细胞的抗增殖作用并促进其凋亡的应用效果。方法选择CCK-8、Annexin-FITC/PI双染色法对紫杉醇抑制非小细胞肺癌的细胞株A549、H460细胞周期作用进行检测,并选择实时荧光定量PCR (qRT-PCR)法对经紫杉醇处理、未经紫杉醇处理(空白对照组)后A549、H460细胞中不同miRNA的表达情况进行检测。结果紫杉醇可对A549、H460细胞增殖进行明显抑制,且随着应用剂量增加,A549、H460细胞的活性不断降低,显示出时间依赖性;经紫杉醇刺激培养后,A549、H460细胞凋亡水平呈明显升高;A549、H460细胞经紫杉醇刺激后,miR-7相关表达水平明显上调; miR-7过表达后Bcl-2相关mRNA的表达水平明显减少;且miR-7过表达后Bcl-2表达水平明显减少;miR-7 inhibitor可对miR-7表达进行抑制。结论紫杉醇能经诱导细胞凋亡对非小细胞肺癌细胞增殖进行抑制,而经紫杉醇处理后A549、H460细胞系miR-7的表达水平均明显上调,且miR-7过表达可使紫杉醇对非小细胞肺癌细胞相关抗增殖作用、促凋亡作用增强。
Objective To investigate the anti-proliferative and apoptosis-promoting effect of Paclitaxel on nonsmall cell lung cancer(NSCLC) cells by up-regulating miR-7. Methods CCK-8 and Annexin-FITC/PI double staining were used to determine the inhibiting effect of Paclitaxel on the cell cycle of NSCLC cell lines A549 and H460.Real-time quantitative fluorescent PCR(qRT-PCR) was used to examine the expression of different miRNAs in A549 and H460 cells treated with Paclitaxel and without Paclitaxel(blank control group). Results Paclitaxel significantly inhibited the proliferation of A549 and H460 cells, and the activity of A549 and H460 cells decreased along with the increasing dosage, showing time dependence. After Paclitaxel-stimulated culture, the apoptosis levels of A549 and H460 cells increased significantly. After A549 and H460 cells were stimulated by Paclitaxel, the expression level of miR-7 was significantly up-regulated. The expression level of Bcl-2 related mRNA significantly decreased after miR-7overexpression. The expression level of Bcl-2 significantly decreased after miR-7 overexpression. The miR-7 inhibitor could inhibit the expression of miR-7. Conclusion Paclitaxel may inhibit the proliferation of NSCLC cells by inducing apoptosis. The expression level of miR-7 in NSCLC cell lines A549 and H460 is significantly up-regulated after Paclitaxel treatment, and overexpression of miR-7 may enhance the anti-proliferative and pro-apoptotic effects of Paclitaxel on NSCLC cells.
Objective To investigate the anti-proliferative and apoptosis-promoting effect of Paclitaxel on nonsmall cell lung cancer(NSCLC) cells by up-regulating miR-7. Methods CCK-8 and Annexin-FITC/PI double staining were used to determine the inhibiting effect of Paclitaxel on the cell cycle of NSCLC cell lines A549 and H460.Real-time quantitative fluorescent PCR(qRT-PCR) was used to examine the expression of different miRNAs in A549 and H460 cells treated with Paclitaxel and without Paclitaxel(blank control group). Results Paclitaxel significantly inhibited the proliferation of A549 and H460 cells, and the activity of A549 and H460 cells decreased along with the increasing dosage, showing time dependence. After Paclitaxel-stimulated culture, the apoptosis levels of A549 and H460 cells increased significantly. After A549 and H460 cells were stimulated by Paclitaxel, the expression level of miR-7 was significantly up-regulated. The expression level of Bcl-2 related mRNA significantly decreased after miR-7overexpression. The expression level of Bcl-2 significantly decreased after miR-7 overexpression. The miR-7 inhibitor could inhibit the expression of miR-7. Conclusion Paclitaxel may inhibit the proliferation of NSCLC cells by inducing apoptosis. The expression level of miR-7 in NSCLC cell lines A549 and H460 is significantly up-regulated after Paclitaxel treatment, and overexpression of miR-7 may enhance the anti-proliferative and pro-apoptotic effects of Paclitaxel on NSCLC cells.
引文
[1]金军,谢玲.紫杉醇新辅助化疗对非小细胞肺癌患者的疗效及组织IF-ITM3、MMP-9表达的影响[J].医学临床研究,2016,33(7):1322-1324.
[2]郭萌萌,廖珍媛,赵娟娟,等.自身启动子调控miR-7真核表达载体的构建及其意义[J].遵义医学院学报,2015,38(3):219-225.
[3]张连美,仲纪祥,孙苏安.EGFR、ALK和Ki-67在非小细胞肺癌中的表达及相关性分析[J].临床肺科杂志,2017,22(1):4-7.
[4]Hayashi Y,Kuriyama H,Umezu H,et al.ClassⅢbeta-tubulinexpression inl tumor cell is correlated with resistance to docetaxelin patients with completely resected non-small cell lung cancer[J].Intern Med,2014,48(4):203-208.
[5]孔倩,王心悦,蒋日成,等.不同二线方案治疗178例晚期非小细胞肺癌的预后及影响因素[J].中华肿瘤杂志,2016,38(4):294-299.
[6]唐曦,徐炎华,汪春林,等.洛铂胸腔灌注治疗晚期非小细胞肺癌患者恶性胸腔积液的临床观察[J].肿瘤研究与临床,2016,28(11):764-766.
[7]刘佳,任宏,吕卫东,等.STMN 1m RNA在非小细胞肺癌中的表达与紫杉醇类药物化疗效果和患者生存率的关系[J].现代肿瘤医学,2016,24(19):3066-3069.
[8]Liu D,Wu J,Shi GY,et al.Role of XRCC1 and ERCC5 polymorphisms on clinical outcomes in advanced non-small cell lung cancer[J].Genet Mol Res,2014,13(2):3100-3107.
[9]涂启敏,任丹.血清S100A2及S100A6检测对非小细胞肺癌的诊断价值[J].肿瘤研究与临床,2016,28(10):664-668.
[10]张金忠,牟坤,王继松.紫杉醇、顺铂联合重组人血管内皮抑制素对非小细胞肺癌患者的疗效及相关指标的影响[J].中国药房,2016,27(30):4218-4220.
[11]金军,谢玲.紫杉醇新辅助化疗对非小细胞肺癌患者的疗效及组织IF-ITM3、MMP-9表达的影响[J].医学临床研究,2016,33(7):1322-1324.
[12]郑鹏凤,韩福才,郭沁香,等.SPARC及白蛋白结合型紫杉醇在非小细胞肺癌中的进展[J].中华临床医师杂志:电子版,2016,40(14):2136-2140.
[13]张亚琛,梁迪,靳晶,等.非小细胞肺癌相关lncRNA的研究进展[J].中国肺癌杂志,2018,21(1):43-49.
[14]邵扣凤,刘燕文.不同剂型紫杉醇联合顺铂治疗晚期非小细胞肺癌的效果分析[J].临床肿瘤学杂志,2016,21(10):893-897.
[15]刘朝富,林益坤,符剑鸣,等.非小细胞肺癌中微RNA的功能及临床价值的研究进展[J].医学综述,2018,24(4):693-697.
[2]郭萌萌,廖珍媛,赵娟娟,等.自身启动子调控miR-7真核表达载体的构建及其意义[J].遵义医学院学报,2015,38(3):219-225.
[3]张连美,仲纪祥,孙苏安.EGFR、ALK和Ki-67在非小细胞肺癌中的表达及相关性分析[J].临床肺科杂志,2017,22(1):4-7.
[4]Hayashi Y,Kuriyama H,Umezu H,et al.ClassⅢbeta-tubulinexpression inl tumor cell is correlated with resistance to docetaxelin patients with completely resected non-small cell lung cancer[J].Intern Med,2014,48(4):203-208.
[5]孔倩,王心悦,蒋日成,等.不同二线方案治疗178例晚期非小细胞肺癌的预后及影响因素[J].中华肿瘤杂志,2016,38(4):294-299.
[6]唐曦,徐炎华,汪春林,等.洛铂胸腔灌注治疗晚期非小细胞肺癌患者恶性胸腔积液的临床观察[J].肿瘤研究与临床,2016,28(11):764-766.
[7]刘佳,任宏,吕卫东,等.STMN 1m RNA在非小细胞肺癌中的表达与紫杉醇类药物化疗效果和患者生存率的关系[J].现代肿瘤医学,2016,24(19):3066-3069.
[8]Liu D,Wu J,Shi GY,et al.Role of XRCC1 and ERCC5 polymorphisms on clinical outcomes in advanced non-small cell lung cancer[J].Genet Mol Res,2014,13(2):3100-3107.
[9]涂启敏,任丹.血清S100A2及S100A6检测对非小细胞肺癌的诊断价值[J].肿瘤研究与临床,2016,28(10):664-668.
[10]张金忠,牟坤,王继松.紫杉醇、顺铂联合重组人血管内皮抑制素对非小细胞肺癌患者的疗效及相关指标的影响[J].中国药房,2016,27(30):4218-4220.
[11]金军,谢玲.紫杉醇新辅助化疗对非小细胞肺癌患者的疗效及组织IF-ITM3、MMP-9表达的影响[J].医学临床研究,2016,33(7):1322-1324.
[12]郑鹏凤,韩福才,郭沁香,等.SPARC及白蛋白结合型紫杉醇在非小细胞肺癌中的进展[J].中华临床医师杂志:电子版,2016,40(14):2136-2140.
[13]张亚琛,梁迪,靳晶,等.非小细胞肺癌相关lncRNA的研究进展[J].中国肺癌杂志,2018,21(1):43-49.
[14]邵扣凤,刘燕文.不同剂型紫杉醇联合顺铂治疗晚期非小细胞肺癌的效果分析[J].临床肿瘤学杂志,2016,21(10):893-897.
[15]刘朝富,林益坤,符剑鸣,等.非小细胞肺癌中微RNA的功能及临床价值的研究进展[J].医学综述,2018,24(4):693-697.