粉防己碱调节cGAS-STING信号通路对黑色素瘤细胞增殖、凋亡和免疫逃逸的影响  

Influences of tetrandrine on proliferation,apoptosis and immune escape of melanoma cells by regulating cGAS-STING signal pathway

作　　者：周慧 李婕[1] 胡利娟 刘慧梅 张伟[2] 王丽娜 ZHOU Hui;LI Jie;HU Lijuan;LIU Huimei;ZHANG Wei;WANG Lina(Pharmacy Department,Qingdao Hospital of Traditional Chinese Medicine(Qingdao Hiser Hospital),Qingdao 266033,China;Skin Department,Qingdao Hospital of Traditional Chinese Medicine(Qingdao Hiser Hospital),Qingdao 266033,China)

机构地区：[1]青岛市中医医院(青岛市海慈医院)药剂科,青岛266033 [2]青岛市中医医院(青岛市海慈医院)皮肤科,青岛266033

出　　处：《中国免疫学杂志》2025年第2期357-361,共5页Chinese Journal of Immunology

基　　金：山东省中医药科技项目(2020M109)。

摘　　要：目的:探讨粉防己碱(Tet)调节环磷酸鸟苷-腺苷酸合成酶(cGAS)-干扰素基因刺激因子(STING)信号通路对黑色素瘤细胞增殖、凋亡和免疫逃逸的影响。方法:将MV3细胞分为NC组、Tet-L组(5μmol/L)、Tet-M组(10μmol/L)、Tet-H组(15μmol/L)、RU.521(cGAS抑制剂)组(1μmol/L)、Tet-H+RU.521组(15μmol/L+1μmol/L),CCK-8和平板克隆实验检测MV3细胞增殖;流式细胞术检测MV3细胞凋亡;Western blot检测MV3细胞增殖细胞核抗原(PCNA)、Bcl-2相关X蛋白(Bax)、TGF-β、cGAS、STING蛋白表达。Transwell小室将各组细胞分别与NK细胞共培养48 h,并命名为NC共培养组、Tet-L共培养组、Tet-M共培养组、Tet-H共培养组、RU.521共培养组、Tet-H+RU.521共培养组,ELISA检测共培养细胞上清中IFN-γ、粒酶B(Granzyme B)水平;检测NK细胞杀伤活性变化。结果:与NC组比较,Tet-L组、Tet-M组、Tet-H组MV3细胞A450、克隆形成率、PCNA、TGF-β蛋白表达降低,细胞凋亡率、Bax、cGAS、STING蛋白表达升高,且呈剂量依赖性,RU.521组对应指标变化趋势与上述相反(P<0.05);与NC共培养组比较,Tet-L共培养组、Tet-M共培养组、Tet-H共培养组细胞上清中IFN-γ、Granzyme B水平、NK细胞杀伤活性升高,且呈剂量依赖性,RU.521共培养组对应指标变化趋势与上述相反(P<0.05);RU.521减弱了高剂量Tet对MV3细胞增殖、免疫逃逸的抑制作用以及对细胞凋亡的促进作用。结论:Tet可能通过激活cGAS-STING信号通路抑制黑色素瘤细胞增殖、免疫逃逸,促进细胞凋亡。Objective:To investigate influences of tetrandrine(Tet)on proliferation,apoptosis and immune escape of melanoma cells by regulating cyclic GMP-AMP synthase(cGAS)-stimulator of interferon gene(STING)signal pathway.Methods:MV3 cells were divided into NC group,Tet-L group(5µmol/L),Tet-M group(10µmol/L),Tet-H group(15µmol/L),RU.521(cGAS inhibitor)group(1µmol/L),Tet-H+RU.521 group(15µmol/L+1µmol/L).Proliferation of MV3 cells was detected by CCK-8 and plate cloning test;apoptosis of MV3 cells was detected by flow cytometry;Western blot was used to detect expressions of proliferating cell nuclear antigen(PCNA),Bcl-2-associated X protein(Bax),TGF-β,cGAS,STING proteins in MV3 cells.Cells in above groups were co-cultured with NK cells for 48 h through Transwell chamber,and named NC co-culture group,Tet-L co-culture group,Tet-M co-culture group,Tet-H co-culture group,RU.521 co-culture group,Tet-H+RU.521 co-culture group,respectively.ELISA was used to detect levels of IFN-γand Granzyme B in supernatant of co-cultured cells;change of NK cell killing activity was detected.Results:Compared with NC group,A450,cloning rate,PCNA and TGF-βprotein expressions of MV3 cells in Tet-L group,Tet-M group and Tet-H group were decreased,apoptosis rate,Bax,cGAS,STING protein expressions were increased,which was in a dose dependent manner,change trend of corresponding indexes in RU.521 group was contrary to the above(P<0.05);compared with NC co-culture group,levels of IFN-γ,Granzyme B and NK cell killing activity in supernatant of Tet-L co-culture group,Tet-M co-culture group and Tet-H co-culture group were increased,which was in a dose-dependent manner,change trend of corresponding indexes in RU.521 co-culture group was opposite to the above(P<0.05);RU.521 attenuated inhibition of high-dose Tet on MV3 cell proliferation,immune escape and its promotion on apoptosis.Conclusion:Tet may inhibit melanoma cell proliferation,immune escape and promote cell apoptosis by activating cGAS-STING signal pathway.

关 键 词：粉防己碱 cGAS-STING信号通路 黑色素瘤 增殖 凋亡 免疫逃逸 

分 类 号：R73-37[医药卫生—肿瘤]