野芋提取物通过SFTA1P对肝癌细胞生物学行为的影响

Effect of Colocasia Antiquorum Extract on Biological Behaviors of Liver Cancer Cells via SFTA1P

目的探讨野芋提取物对肝癌细胞生物行为的影响及分子机制。方法用浓度分别为5、10、15、20 mg/L的野芋提取物处理HCC9204肝癌细胞,作为不同剂量野芋提取物组;正常培养的HCC9204细胞作为对照组;将pcDNA3.1、pcDNA3.1-SFTA1P转染至HCC9204细胞中,记为pcDNA3.1组、pcDNA3.1-SFTA1P组;将si-NC、si-SFTA1P转染至HCC9204细胞后用20 mg/L的野芋提取物处理,记为野芋提取物+si-NC组,野芋提取物+si-SFTA1P组。细胞计数试剂盒8(CCK-8)检测细胞存活率;流式细胞术检测细胞凋亡;克隆形成实验检测细胞增殖;Transwell实验检测细胞迁移;实时荧光定量PCR(qRT-PCR)检测SFTA1P和miR-665的表达水平;双荧光素酶报告实验检测SFTA1P和miR-665的靶向关系。结果野芋提取物处理的肝癌HCC9204细胞,细胞存活率显著降低,细胞凋亡率显著升高,克隆形成数显著减少,迁移细胞数显著减少(均P<0.05)。过表达SFTA1P可抑制肝癌HCC9204细胞存活、迁移,促进细胞凋亡。SFTA1P靶向调控miR-665。抑制SFTA1P表达可逆转野芋提取物对HCC9204细胞增殖、迁移和凋亡的影响。结论野芋提取物可通过调控SFTA1P/miR-665轴抑制肝癌HCC9204细胞增殖、迁移,促进细胞凋亡。

Objective To investigate the effect of Colocasia Antiquorum extract on the biological behaviors of liver cancer cells and molecular mechanism.Methods HCC9204 cells treated with 5,10,15,and 20 mg/L Colocasia Antiquorum extracts were set as different doses of Colocasia Antiquorum extract groups respectively.Normal cultured cells were used as control group.pcDNA3.1 and pcDNA3.1-SFTA1P were transfected into HCC9204 cells and recorded as pcDNA3.1 group and pcDNA3.1-SFTA1P group.HCC9204 cells transfected with si-NC and si-SFTA1P were treated with 20 mg/L Colocasia Antiquorum extract and recorded as Colocasia Antiquorum extract+si-NC group,Colocasia Antiquorum extract+si-SFTA1P group respectively.Cell counting kit 8(CCK-8)was used to detect cell survival rate.Flow cytometry was used to detect cell apoptosis.Colony formation assay was used to detect the number of cell clones.Transwell assay was used to detect cell migration.Real-time quantitative polymerase chain reaction(qRT-PCR)was used to detect the expression levels of SFTA1P and miR-665.Dual luciferase reporter assay was used to detect the targeting relationship between SFTA1P and miR-665.Results In liver cancer HCC9204 cells treated with Colocasia Antiquorum extract,the cell survival rate was significantly reduced,the apoptosis rate was significantly increased,the numbers of colony-forming cells and migrated cells were significantly reduced(all P<0.05).Overexpression of SFTA1P could inhibit the survival and migration of HCC9204 cells yet promote cell apoptosis.SFTA1P could target miR-665.Inhibition of SFTA1P expression could reverse the effects of Colocasia Antiquorum extract on the proliferation,migration and apoptosis of HCC9204 cells.Conclusion Colocasia Antiquorum extract can inhibit the proliferation,migration while promote the apoptosis of HCC9204 cells by regulating the SFTA1P/miR-665 axis.