川陈皮素通过诱导细胞自噬抑制宫颈癌细胞侵袭能力

Effect of nobiletin inhibiting human cervical carcinoma cells invasion by inducing autophagy

目的了解川陈皮素(nobiletin)对宫颈癌Si Ha细胞侵袭能力及自噬水平的影响。方法选择宫颈癌SiHa细胞作为实验对象,用不同浓度的川陈皮素(0、25、50、100 mg/L)在体外处理SiHa细胞,Transwell小室法检测细胞侵袭能力的改变;蛋白质印迹法检测细胞EMT标志物(E-cadherin、Vimentin、Fibronectin)蛋白、自噬相关基因蛋白(Atg5、LC3-II和Beclin1蛋白)和金属基质蛋白酶9(MMP-9)蛋白表达变化情况。结果 Transwell侵袭实验结果显示,川陈皮素处理24 h后可抑制宫颈癌SiHa细胞侵袭能力,25、50、100 mg/L川陈皮素组的穿膜细胞数分别为(318.6±12.9)、(221.5±11.3)和(109.8±13.7)个,与对照组的穿膜细胞数[(423.4±16.6)个]比较,差异有统计学意义(P<0.05);蛋白质印迹结果显示,川陈皮素可促进Si Ha细胞EMT标志物蛋白E-cadherin的表达,同时抑制Vimentin、Fibronectin蛋白的表达,且抑制MMP-9蛋白表达;川陈皮素也可上调自噬相关蛋白Atg5、LC3-II和Beclin1的表达。结论川陈皮素可抑制宫颈癌SiHa细胞侵袭能力,其机制可能通过诱导Si Ha细胞自噬并抑制EMT形成来实现。

Objective To explore the effects of nobiletin on the metastasis and autophagy in human cervical carcinoma SiHa cells.Methods The cervical carcinoma SiHa cells were cultured,and were treated with nobiletin at different concentrations(0,25,50,100 mg/L)in vitro.Transwell chamber assay was used to detect the changes in cell invasiveness.Western blotting were used to test the expression changes in EMT markers(E-cadherin,Vimentin,Fibronectin)protein,autophagy-related gene protein(Atg5,LC3-II and Beclin1 protein)and metal matrix protease 9(MMP-9)protein.Results After treated with nobiletin(25,50 and 100 mg/L),the number of transwell cells was 318.6±12.9,221.5±11.3 and 109.8±13.7 respectively,and the number of SiHa cells treated by nobiletin passing through the basement membrane was less than that of the control group(423.4±16.6),and the parameters showed a significant difference(P<0.05),suggesting that nobiletin could inhibit the capacity of cell invasion.Nobiletin could promoted the expressions of E-cadherin,Atg5,LC3-II and Beclin1 protein,and suppressed the expressions of Vimentin,Fibronectin,MMP-9.Conclusion Nobiletin could inhibit the capacity of cell invasion,which may be associated with promotion of autophagy and inhibition of epithelial-to-mesenchymal transition in cervical cancer cell line SiHa cells.