| 邹华兰,曾健梅,冷兴强,等.葛根素联合 γ突触核蛋白 -微小 RNA对肺癌 H1299细胞增殖凋亡的影响[J].安徽医药,2021,25(5):1009-1013. |
| 葛根素联合 γ突触核蛋白 -微小 RNA对肺癌 H1299细胞增殖凋亡的影响 |
| Effect of puerarin combined with SNCG-siRNA on proliferation and apoptosis of lung cancer H1299 cells and its mechanism |
| |
| DOI：10.3969/j.issn.1009-6469.2021.05.041 |
| 中文关键词: 肺肿瘤 葛根素 γ突触核蛋白 细胞增殖 细胞凋亡 |
| 英文关键词: Lung neoplasms Puerarin SNCG Cell proliferation Cell apoptosis |
| 基金项目: |
|
| 摘要点击次数: 1260 |
| 全文下载次数: 401 |
| 中文摘要: |
| 目的探讨葛根素联合 γ突触核蛋白 -微小 RNA(SNCG-siRNA)对肺癌 H1299细胞增殖、凋亡的影响及其机制。方法以 0、20、40、80、160和 320 μg/mL葛根素作用于体外培养的 H1299细胞 24 h后，采用 MTT法检测细胞增殖并计算出葛根素的 IC50。在 H1299细胞中转染 SNCG干扰序列 SNCG-siRNA及其阴性对照 NC-siRNA后，采用逆转录 -聚合酶链反应( RT-PCR)和蛋白质免疫印迹(Western blot)检测转染效果。以 150 μg/mL葛根素处理转染 SNCG-siRNA或 NC-siRNA的 H1299细胞 24 h后，采用 MTT法、克隆形成实验和流式细胞仪分别检测细胞的增殖、克隆形成和凋亡能力， Western blot检测细胞中肿瘤增殖抗原(Ki67)、细胞周期蛋白 D1(Cyclin D1)、 B淋巴细胞瘤 -2(Bcl-2)蛋白和 Bcl-2相关 X蛋白( Bax)的表达情况。结果与 0 μg/mL处理组相比， 20、40、80、160和 320 μg/mL葛根素处理后 H1299细胞存活率均明显降低( P<0.05)，IC50为 150.40 μg/mL。与 NCsiRNA组相比， SNCG-siRNA组细胞中 SNCG mRNA和蛋白的表达水平均明显降低( P<0.05)。与 NC-siRNA组相比， SNCG-siRNA、葛根素 +NC-siRNA和葛根素 +SNCG-siRNA处理组细胞的存活率、克隆形成率和细胞中 Ki67、CyclinD1、Bcl-2蛋白的表达水平均明显降低，而细胞凋亡率和细胞中 Bax蛋白的表达水平均明显升高( P<0.05)，且葛根素 +SNCG-siRNA处理对 H1299细胞的作用强度明显大于葛根素 +NC-siRNA或者 SNCG-siRNA组。结论葛根素素和 SNCG-siRNA联合可协同抑制 H1299细胞增殖并促进细胞凋亡，其作用机制可能与共同下调 Ki67、CyclinD1、Bcl-2和上调 Bax蛋白的表达有关。 |
| 英文摘要: |
| Objective To investigate the effect of puerarin combined with SNCG-siRNA on proliferation and apoptosis of lung cancer H1299 cells and its mechanism.Methods Puerarin (0, 20, 40, 80, 160 and 320 μg/mL) was used to treat H1299 cells in vitro for 24 hours. MTT method was used to detect cell proliferation and calculate the IC50 of puerarin. After transfection of SNCG interferencesequence SNCG-siRNA and its negative control NC-siRNA into H1299 cells, the transfection effect was detected by RT-PCR and Western blot. The proliferation, clone formation and apoptosis of H1299 cells transfected with SNCG-siRNA or NC-siRNA were detected byMTT, clone formation test and flow cytometry after treatment with 150μg/mL puerarin for 24 hours. The expressions of Ki67, Cyclin D1,Bax and Bcl-2 protein were detected by Western blot.Results Compared with 0μg/mL treatment group, the survival rate of H1299cells stimulated by 20, 40, 80, 160 and 320 μg/mL puerarin was significantly decreased (P<0.05), and the IC50 was 150.40 μg/mL. Compared with NC-siRNA group, the expression levels of SNCG mRNA and protein in SNCG-siRNA group were significantly lower (P< 0.05). Compared with NC-siRNA group, the cell survival rate, cloning formation rate and the expression level of Ki67, Cyclin D1 and Bcl-2 protein were reduced significantly in SNCG-siRNA, puerarin + NC-siRNA and puerarin + SNCG-siRNA treatment groups, while the apoptotic rate and the expression level of Bax protein in cells were significantly increased (P<0.05), and the effect of puerarin + SNCG-siRNA treatment on H1299 cells was stronger than that of puerarin + NC-siRNA or SNCG-siRNA.Conclusion Puerarin and SNCG-siRNA can synergistically inhibit the proliferation and promote apoptosis of H1299 cells. The mechanism may be related to the co-down-regulation of Ki67, Cyclin D1, Bcl-2 and up-regulation of Bax protein expression. |
|
查看全文
查看/发表评论 下载PDF阅读器 |
| 关闭 |
|
|
|
