香加皮宝藿苷-Ⅰ抑制结肠癌细胞株SW480和RKO的恶性表型及其作用机制

Baohuoside-Ⅰ inhibits malignant phenotype of SW480 and RKO cells and its mechanism

目的:探讨香加皮宝藿苷-Ⅰ对人结肠癌细胞株SW480和RKO增殖、侵袭、迁移和凋亡的影响,并对其机制进行初步的探讨。方法:用不同质量浓度(0、5、10、20μg/ml)的香加皮宝藿苷-Ⅰ溶液分别处理结肠癌细胞株SW480和RKO,采用MTT法检测细胞的增殖情况,通过细胞克隆形成实验检测细胞克隆形成能力,通过Transwell方法检测细胞的迁移和侵袭能力,通过流式细胞仪检测细胞凋亡和细胞周期。通过Western blotting检测C-PARP、Bcl-2与caspase-3蛋白的表达水平,通过RNA-seq检测分析香加皮宝藿苷-Ⅰ对SW480细胞转录组的影响及其可能影响的信号通路。结果:香加皮宝藿苷-Ⅰ能抑制SW480和RKO细胞的增殖、侵袭和迁移,并且能够诱导细胞凋亡和阻滞于细胞G0/G1期。香加皮宝藿苷-Ⅰ处理可上调SW480和RKO细胞株中C-PARP与caspase-3蛋白的表达水平、下调Bcl-2蛋白的表达水平;RNA-seq数据分析显示,SW480细胞DNA复制及ERBB信号通路等相关基因的转录都受到香加皮宝藿苷-Ⅰ的影响。结论:香加皮宝藿苷-Ⅰ通过影响凋亡相关蛋白的表达、细胞DNA复制和ERBB信号通路来诱导细胞凋亡和细胞G0/G1期阻滞,进而抑制人结肠癌细胞株SW480和RKO的恶性表型。

Objective: To investigate the effect of Baohuoside-Ⅰ on the proliferation, invasion, migration and apoptosis of colon cancer cell lines SW480 and RKO and the relative mechanism. Methods: Colon cancer cell lines SW480 and RKO were respectively treated with different concentrations of Baohuoside-Ⅰ(0, 5, 10, 20 μg/ml). Cell proliferation was detected by MTT assay;The ability of cell clone formation was tested by cell clone formation experiments;The migration and invasion of cells were detected by Transwell assay;The apoptosis and cell cycle was detected by Flow cytometry;and the protein expression levels of cleaved PARP, cleaved Caspase-3 and Bcl-2 were detected by Western blotting. The effects of Baohuoside-Ⅰ on transcriptome and possible signaling pathways were detected by RNA-Seq technology. Results: Baohuoside-Ⅰ could inhibit the proliferation, invasion and migration of SW480 and RKO cells, and induce cell apoptosis and G0/G1 phase block. Baohuoside-Ⅰ could also up-regulate the protein expressions of cleaved PARP and cleaved Caspase-3 but down-regulate the protein expression of Bcl-2 in SW480 and RKO cell lines. In addition, RNA-Seq data analysis showed that DNA replication and transcription of ERBB signaling pathway related genes were both affected by Baohuoside-Ⅰ. Conclusion: Baohuoside-Ⅰ could induce apoptosis and G0/G1 phase block of colon cancer cell lines SW480 and RKO by affecting the expression of apoptosis related proteins, as well as cellular DNA replication and ERBB signaling pathways, thus inhibiting the malignant phenotypes of SW480 and RKO.