摘要
目的:探讨干扰素-γ(INF-γ)对黑色素瘤细胞的影响及作用机制。方法:选取人黑色素瘤细胞A375,分为对照组(空白未处理)、INF-γ低剂量组(加入INF-γ10μg/L)、中剂量组(加入INF-γ50μg/L)、高剂量组(加入INF-γ100μg/L)。采用CCK-8法检测A375细胞存活情况。采用流式细胞术检测A375细胞凋亡情况。采用Transwell实验检测A375细胞侵袭能力。采用细胞划痕实验检测A375细胞迁移能力。采用Western blot检测A375细胞磷脂酰肌醇3-激酶(PI3K)、p-PI3K、蛋白激酶B(AKT)、p-AKT、哺乳动物雷帕霉素靶蛋白(mTOR)蛋白表达。根据细胞分组,将BALB/c裸鼠也相应随机分为4组,每组8只,建立裸鼠皮下移植瘤模型。比较各组裸鼠不同时间皮下移植瘤体积。结果:与对照组比较,INF-γ低、中、高剂量组A375细胞存活率(24、48、72 h)、迁移率、细胞侵袭数、裸鼠皮下移植瘤体积(7、14、21、28 d)以及p-PI3K、p-AKT、mTOR蛋白表达水平依次降低,细胞凋亡率依次升高(均P<0.05)。结论:INF-γ可通过下调PI3K/AKT/mTOR信号通路抑制黑色素瘤细胞侵袭、迁移以及裸鼠皮下成瘤速度,并诱导细胞凋亡。
Objective:To explore the effects of interferon-γ(IFN-γ)on melanoma cells and its mechanisms.Methods:Human melanoma cells A375 were selected and divided into a control group(untreated),a low-dose IFN-γgroup(with 10μg/L IFN-γ),a medium-dose group(with 50μg/L IFN-γ),and a high-dose group(with 100μg/L IFN-γ).The CCK-8 assay and flow cytometry were used to detect the survival and apoptosis of A375 cells.The Transwell assay and scratch test were employed to assess the invasive and migratory capabilities of A375 cells.Western blot was utilized to measure the protein expression of PI3K,p-PI3K,AKT,p-AKT and mTOR in A375 cells.BALB/c nude mice were also randomly divided into four groups,with each group containing eight mice,to establish a nude mouse subcutaneous xenograft model.The subcutaneous tumor volumes of nude mice at different time points were compared among the groups.Results:Compared to the control group,the survival rate of A375 cells(at 24,48,and 72 hours),migration rate,number of invasive cells,subcutaneous tumor volume in nude mice(at 7,14,21,and 28 days),and the expression levels of p-PI3K,p-AKT,and mTOR proteins were progressively decreased with increasing doses of IFN-γ,while the apoptosis rate was progressively increased(all P<0.05).Conclusion:IFN-γcan inhibit the invasion,migration and subcutaneous tumor growth rate of melanoma cells in nude mice,and induce cell apoptosis by down-regulating the PI3K/AKT/mTOR signaling pathway.
作者
张梦迪
孙琼
拓惠惠
巨曼钰
郑焱
ZHANG Mengdi;SUN Qiong;TUO Huihui;JU Manyu;ZHENG Yan(Department of Dermatology,the Second Affiliated Hospital of Xi’an Jiaotong University,Xi’an 710004,China)
出处
《陕西医学杂志》
CAS
2024年第11期1459-1462,1467,共5页
Shaanxi Medical Journal
基金
国家自然科学基金资助项目(81972938)。
