四逆汤协同壮观霉素B1诱导PTEN与SUMO1解离治疗肺癌的研究

Sini Decoction combined with spectinomycin B1 induced dissociation of PTEN and SUMO1 to treat lung cancer

目的从蛋白质类泛素化修饰角度解析温里剂四逆汤协同壮观霉素B1抑制肺癌细胞增殖、迁移和促进细胞凋亡的作用机制。方法实验分对照组、壮观霉素B1组、四逆汤组和联合药物组,于人非小细胞肺癌A549细胞株作用48 h后,采用Westernblotting法检测小泛素相关修饰蛋白-1(SUMO1)、10号染色体缺失磷酸酶和张力蛋白同源物基因(PTEN)、磷酸化Akt(p-AKT)和Caspase-9的蛋白表达水平;MTT实验绘制细胞增殖曲线;流式细胞术检测细胞凋亡;细胞划痕和Transwell实验检测细胞迁移和侵袭。结果壮观霉素B1和四逆汤均能够降低A549细胞共价态SUMO1和p-Akt水平,增加PTEN和活化Caspase-9的蛋白水平,抑制细胞增殖、迁移和侵袭,促进细胞凋亡,壮观霉素B1和四逆汤联合药物组对细胞的抑制作用显著强于各单独给药组。结论四逆汤能够协同壮观霉素B1诱导PTEN与SUMO1解离,进而抑制肺癌细胞增殖、转移和促进细胞凋亡。

Objective To analyze the mechanism of the inhibition of proliferation, metastasis and the promotion of apoptosis of lung cancer cells by Sini Decoction combined with spectinomycin B1 from the perspective of protein SUMOylation modification.Methods The experiment was divided into control group, spectinomycin B1 group, Sini Decoction group, and combined treatment group. Human non-small cell lung cancer cell line A549 was treated for 48 h and Western blotting was used to detect the protein expression of small ubiquitin-related modified protein-1(SUMO1), gene of phosphate and tension homology deleted on chromsome ten(PTEN), and phosphorylated Akt(p-Akt) and Caspase-9. Cell proliferation curves were drawn by MTT assay, and cell apoptosis was detected by flow cytometry;Cell migration and cell invasion were detected by cell scratch and Transwell assay. Results Both spectinomycin B1 and Sini Decoction an reduce the levels of covalent SUMO1 and p-Akt, increase the protein levels of PTEN and activate Caspase-9, inhibit cell proliferation, migration and invasion, and promote cell apoptosis. Combined treatment group can further enhance the above changes. Conclusion Sini Decoction can synergize with spectinomycin B1 to induce the dissociation of PTEN and SUMO1, thereby inhibiting the proliferation, metastasis and promoting apoptosis of lung cancer cells.