摘要
目的探讨蛇床子素(osthole或osthol)能否通过PI3K/Akt通路诱导细胞凋亡,抑制细胞增殖。方法体外培养外阴鳞癌SW962细胞,加入不同浓度蛇床子素,MTT法检测细胞存活情况,DAPI染色观察细胞形态学变化;流式细胞术检测SW962细胞凋亡情况; Western blot检测Bcl-2、Bax、cleaved caspase-3、PI3K和p-Akt蛋白表达,及蛇床子素和IGF-1共作用下PI3K/Akt通路蛋白的变化。结果 MTT结果显示,蛇床子素呈浓度依赖性地抑制SW962细胞增殖;DAPI染色和流式细胞术结果显示,蛇床子素呈浓度依赖性诱导细胞凋亡。Western blot结果显示,蛇床子素能上调Bax和cleaved caspase-3蛋白表达,下调PI3K、p-Akt和Bcl-2蛋白表达。IGF-1诱导PI3K、p-Akt和Bcl-2蛋白表达增加,蛇床子素逆转IGF-1作用,3种蛋白表达下调。结论蛇床子素通过抑制PI3K/Akt通路,诱导外阴鳞癌SW962细胞凋亡,抑制细胞增殖。
Aim To investigate the effect of osthole on inducing apoptosis and inhibiting the proliferation of vulvar squamous cell carcinoma SW962 cells through PI3 K/Akt signaling pathway and changes of PI3 K/Akt pathway protein in the coexistence of osthole and IGF-1. Methods Vulvar squamous cell carcinoma SW962 cells were cultured in vitro and different concentrations of osthole intervention were given. The effect of osthole on the proliferation of SW962 cells was determined by MTT. Cell apoptosis was detected by flow cytometry.DAPI staining revealed the changes in cellular morphology. The expressions of cleaved-caspase-3,PI3 K,p-Akt,Bcl-2,Bax protein in SW962 cells were assessed by Western blot. Results MTT results showed that osthole could inhibit the proliferation of SW962 cells in a concentration-dependent manner. The results of DAPI staining and FCM by Annexin V-FITC and PI staining indicated osthole induced apoptosis of SW962 cells in a concentration-dependent manner. Western blot showed that osthole increased the expression of Bax and cleaved caspase-3 and decreased the expression of Bcl-2,PI3 K,p-Akt. IGF-1 induced increased expression of PI3 K,p-Akt and Bcl-2,while osthole reversed IGF-1 and down-regulated the expression of three proteins. Conclusions Osthole induces the apoptosis and inhibits the proliferation of SW962 in vulvar squamous cell carcinoma by inhibiting PI3 K/Akt signaling pathway.
引文
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