常春藤皂苷元(hederagenin)通过抑制STAT3通路降低CaSki宫颈癌细胞增殖能力并促进其凋亡
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摘要
目的探讨常春藤皂苷元(hederagenin)对宫颈癌细胞活性及凋亡的影响及机制。方法培养CaSki宫颈癌细胞,采用(0、 10、 20、 40、 80)μg/mL常春藤皂苷元处理,噻唑蓝(MTT)法测定细胞活性,确定其半数抑制剂量约40μg/mL。采用40μg/mL常春藤皂苷元处理宫颈癌细胞,流式细胞术测定宫颈癌细胞凋亡、 Western blot法测定宫颈癌细胞裂解型胱天蛋白酶3(c-caspase-3)、 c-caspase-9、信号转导子和转录激活因子3(STAT3)、磷酸化的STAT3(p-STAT3)蛋白水平。用STAT3信号通路抑制剂AG490和常春藤皂苷元共同处理宫颈癌细胞,用上述方法检测细胞活性和凋亡水平。结果与对照组细胞相比, 40μg/mL常春藤皂苷元处理后的宫颈癌细胞增殖活性降低,细胞凋亡水平升高, c-caspase-3和c-caspase-9水平增加, p-STAT3蛋白水平降低。与常春藤皂苷元处理细胞相比, AG490和常春藤皂苷元共同处理的宫颈癌细胞活性进一步降低,细胞凋亡率增加,细胞中c-caspase-3和c-caspase-9的蛋白水平升高。结论常春藤皂苷元抑制宫颈癌细胞的增殖并诱导其凋亡,与抑制STAT3信号通路有关。
Objective To investigate the effect of hederagenin on the activity and apoptosis of cervical cancer cells and its mechanism. Methods Cervical cancer Ca Ski cells were cultured and treated with 0,10,20,40,80 μg/m L hederagenin.Cell viability was determined by MTT assay. The 50% inhibitory concentration( IC50) was determined to be about 40 μg/m L.The cervical cancer cells were treated with 40 μg/m L hederagenin. The apoptosis of cervical cancer cells was measured by flow cytometry,and the levels of cleaved caspase-3( c-caspase-3),c-caspase-9,and STAT3 and p-STAT3 proteins were determined by Western blot analysis. STAT3 signaling pathway inhibitor AG490 and hederagenin were used to treat cervical cancer cells,and then the activity and apoptotic level of cervical cancer cells were detected by the above methods. Results Compared with the control cells,the proliferation activity of the cervical cancer cells treated with 40 μg/m L hederagenin decreased,the apoptotic level increased,the levels of c-caspase-3 and c-caspase-9 increased,and the level of p-STAT3 protein decreased. Compared with the cells treated with hederagenin,the activity of cervical cancer cells treated with AG490 and hederagenin decreased further,the apoptotic rate increased,and the protein levels of c-caspase-3 and c-caspase-9 increased.Conclusion Hederagenin inhibits the proliferation and promotes the apoptosis of cervical cancer cells,which is related to the inhibition of STAT3 signaling pathway.
Objective To investigate the effect of hederagenin on the activity and apoptosis of cervical cancer cells and its mechanism. Methods Cervical cancer Ca Ski cells were cultured and treated with 0,10,20,40,80 μg/m L hederagenin.Cell viability was determined by MTT assay. The 50% inhibitory concentration( IC50) was determined to be about 40 μg/m L.The cervical cancer cells were treated with 40 μg/m L hederagenin. The apoptosis of cervical cancer cells was measured by flow cytometry,and the levels of cleaved caspase-3( c-caspase-3),c-caspase-9,and STAT3 and p-STAT3 proteins were determined by Western blot analysis. STAT3 signaling pathway inhibitor AG490 and hederagenin were used to treat cervical cancer cells,and then the activity and apoptotic level of cervical cancer cells were detected by the above methods. Results Compared with the control cells,the proliferation activity of the cervical cancer cells treated with 40 μg/m L hederagenin decreased,the apoptotic level increased,the levels of c-caspase-3 and c-caspase-9 increased,and the level of p-STAT3 protein decreased. Compared with the cells treated with hederagenin,the activity of cervical cancer cells treated with AG490 and hederagenin decreased further,the apoptotic rate increased,and the protein levels of c-caspase-3 and c-caspase-9 increased.Conclusion Hederagenin inhibits the proliferation and promotes the apoptosis of cervical cancer cells,which is related to the inhibition of STAT3 signaling pathway.
引文
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[17] Wan L,Zhang D,Zhang J,et al.TT-1,an analog of melittin,triggers apoptosis in human thyroid cancer TT cells via regulating caspase,Bcl-2 and Bax[J].Oncol Lett,2018,15(1):1271-1278.
[18] 李志强,孙洋,万鸿兴,等.过表达N-myc下游调节基因2(NDRG2)抑制直肠癌细胞的增殖和迁移并促进细胞凋亡[J].细胞与分子免疫学杂志,2017,33(1):48-52.Li Z,Sun Y,Wan H,et al.Overexpression of N-myc downstream regulated gene 2 (NDRG2) inhibits proliferation,migration and promotes apoptosis in SW480 rectal cancer cells[J].Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi,2017,33(1):48-52.
[19] 张铁,彭翠平,王永林,等.α-常春藤皂苷抗肿瘤作用机制研究[J].中药新药与临床药理,2015,26(2):175-179.Zhang T,Peng C,Wang Y,et al.Study on the anti-tumor mechanism of α-hederin[J].Zhong Yao Xin Yao Yu Lin Chuang Yao Li,2015,26(2):175-179.
[20] Rong J,Li L,Jing L,et al.JAK2/STAT3 pathway mediates protection of metallothionein against doxorubicin-induced cytotoxicity in mouse cardiomyocytes[J].Int J Toxicol,2016,35(3):317-326.
[21] Ma C,Horlad H,Pan C,et al.STAT3 inhibitor abrogates the expression of PD-1 ligands on lymphoma cell lines[J].J Clin Exp Hematop,2017,57(1):21-25.
[22] Zhao Z,Lv B,Zhang L,et al.miR-202 functions as a tumor suppressor in non-small cell lung cancer by targeting STAT3[J].Mol Med Rep,2017,16(2):2281-2289.
[23] Ren D,Lin B,Zhang X,et al.Maintenance of cancer stemness by miR-196b-5p contributes to chemoresistance of colorectal cancer cells via activating STAT3 signaling pathway[J].Oncotarget,2017,8(30):49807-49823.
[24] Li Y,Cui N,Zheng P S,et al.BMX/Etk promotes cell proliferation and tumorigenicity of cervical cancer cells through PI3K/AKT/mTOR and STAT3 pathways[J].Oncotarget,2017,8(30):49238-49252.
[25] Liu Y,Gong W,Yang Z Y,et al.Quercetin induces protective autophagy and apoptosis through ER stress via the p-STAT3/Bcl-2 axis in ovarian cancer[J].Apoptosis,2017,22(4):544-557.
[2] 刘包欣子,王瑞平,邹玺,等.常春藤皂苷元对结肠癌细胞LoVo增殖、 粘附、 侵袭和迁移能力的影响[J].南京中医药大学学报,2013,29(1):44-47.Liu B,Wang R,Zou X,et al.The effect of hederagenin on the proliferation,adhesion,invasion and migration of human colon cancer cells LoVo[J].Nanjing Zhong Yi Yao Da Xue Xue Bao,2013,29(1):44-47.
[3] 李欣欣,杨中林,杨小林,等.常春藤皂苷元抑制人乳腺癌MCF-7和肺癌A549裸鼠移植瘤生长研究[J].海峡药学,2016,28(5):16-18.Li X,Yang Z,Yang X,et al.Effect of hederagenin on the growth of human breast cancer MCF-7 and lung cancer A549 xenogeafts in nude mice[J].Hai Xia Yi Yao,2016,28(5):16-18.
[4] 陈彦,奚松阳,滕钰浩,等.常春藤皂苷元对肠癌SW480细胞上皮-间质转化(EMT)及侵袭的影响[J].中国实验方剂学杂志,2016,22(12):133-138.Chen Y,Xi S,Teng Y,et al.Effects of hederagenin in suppressing epithelial-mesenchymal transition (EMT) and invasion of colon cancer cells SW480[J].Zhongguo Shi Yan Fang Ji Xue Za Zhi,2016,22(12):133-138.
[5] Lv Q,Hu J X,Li Y J,et al.miR-320a effectively suppresses lung adenocarcinoma cell proliferation and metastasis by regulating STAT3 signals[J].Cancer Biol Ther,2017,18(3):142-151.
[6] Xu Y,Tong Y,Ying J,et al.Chrysin induces cell growth arrest,apoptosis,and ER stress and inhibits the activation of STAT3 through the generation of ROS in bladder cancer cells[J].Oncol Lett,2018,15(6):9117-9125.
[7] Zhang Z,Wang J,Cheng J,et al.Effects of miR-126 on the STAT3 signaling pathway and the regulation of malignant behavior in lung cancer cells[J].Oncol Lett,2018,15(6):8412-8416.
[8] 张崇媛,胡萍,何秋敏,等.枸杞多糖和AG490联用对宫颈癌细胞增殖和凋亡的影响研究[J].肿瘤药学,2017,7(2):183-188.Zhang C,Hu P,He Q,et al.The effects of Lycium barbarum polysaccharides combined with AG490 on the proliferation and apoptosis of cervical cancer cells[J].Zhong Liu Yao Xue,2017,7(2):183-188.
[9] Liu Z,Zheng Q,Chen W,et al.Chemosensitizing effect of Paris Saponin I on Camptothecin and 10-hydroxycamptothecin in lung cancer cells via p38 MAPK,ERK,and AKT signaling pathways[J].Eur J Med Chem,2017,5(125):760-769.
[10] 刘包欣子,王瑞平,邹玺,等.常春藤皂苷元对胃癌细胞MGC-803增殖、 黏附、 侵袭和迁移能力的影响[J].中国实验方剂学杂志,2013,19(4):212-215.Liu B,Wang R,Zou X,et al.Effect of hederagenin on biological behaviors including proliferation,adhesion,invasion and migration capabilities of human gastric cancer cell line[J].Zhongguo Shi Yan Fang Ji Xue Za Zhi,2013,19(4):212-215.
[11] Ng P Y,Chye S M,Ng C h H,et al.Clinacanthus Nutans Hexane extracts induce apoptosis through a caspase-dependent pathway in human cancer cell lines[J].Asian Pac J Cancer Prev,2017,18(4):917-926.
[12]Yao Q,Wang W,Jin J,et al.Synergistic role of caspase-8 and caspase-3 expressions:Prognostic and predictive biomarkers in colorectal cancer[J].Cancer Biomark,2018,21(4):899-908.
[13] Nayak V L,Nagesh N,Ravikumar A,et al.2-Aryl benzimidazole conjugate induced apoptosis in human breast cancer MCF-7 cells through caspase independent pathway[J].Apoptosis,2017,22(1):118-134.
[14] Peng T,Dou Q P.Everolimus inhibits growth of gemcitabine-resistant pancreatic cancer cells via induction of caspase-dependent apoptosis and G2/M Arrest[J].J Cell Biochem,2017,118(9):2722-2730.
[15] Hong H,An J C,de La Cruz J F,et al.Cnidium officinale Makino extract induces apoptosis through activation of caspase-3 and p53 in human liver cancer HepG2 cells[J].Exp Ther Med,2017,14(4):3191-3197.
[16] Jia W,Yu T,An Q,et al.microRNA-423-5p inhibits colon cancer growth by promoting caspase-dependent apoptosis[J].Exp Ther Med,2018,16(2):1225-1231.
[17] Wan L,Zhang D,Zhang J,et al.TT-1,an analog of melittin,triggers apoptosis in human thyroid cancer TT cells via regulating caspase,Bcl-2 and Bax[J].Oncol Lett,2018,15(1):1271-1278.
[18] 李志强,孙洋,万鸿兴,等.过表达N-myc下游调节基因2(NDRG2)抑制直肠癌细胞的增殖和迁移并促进细胞凋亡[J].细胞与分子免疫学杂志,2017,33(1):48-52.Li Z,Sun Y,Wan H,et al.Overexpression of N-myc downstream regulated gene 2 (NDRG2) inhibits proliferation,migration and promotes apoptosis in SW480 rectal cancer cells[J].Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi,2017,33(1):48-52.
[19] 张铁,彭翠平,王永林,等.α-常春藤皂苷抗肿瘤作用机制研究[J].中药新药与临床药理,2015,26(2):175-179.Zhang T,Peng C,Wang Y,et al.Study on the anti-tumor mechanism of α-hederin[J].Zhong Yao Xin Yao Yu Lin Chuang Yao Li,2015,26(2):175-179.
[20] Rong J,Li L,Jing L,et al.JAK2/STAT3 pathway mediates protection of metallothionein against doxorubicin-induced cytotoxicity in mouse cardiomyocytes[J].Int J Toxicol,2016,35(3):317-326.
[21] Ma C,Horlad H,Pan C,et al.STAT3 inhibitor abrogates the expression of PD-1 ligands on lymphoma cell lines[J].J Clin Exp Hematop,2017,57(1):21-25.
[22] Zhao Z,Lv B,Zhang L,et al.miR-202 functions as a tumor suppressor in non-small cell lung cancer by targeting STAT3[J].Mol Med Rep,2017,16(2):2281-2289.
[23] Ren D,Lin B,Zhang X,et al.Maintenance of cancer stemness by miR-196b-5p contributes to chemoresistance of colorectal cancer cells via activating STAT3 signaling pathway[J].Oncotarget,2017,8(30):49807-49823.
[24] Li Y,Cui N,Zheng P S,et al.BMX/Etk promotes cell proliferation and tumorigenicity of cervical cancer cells through PI3K/AKT/mTOR and STAT3 pathways[J].Oncotarget,2017,8(30):49238-49252.
[25] Liu Y,Gong W,Yang Z Y,et al.Quercetin induces protective autophagy and apoptosis through ER stress via the p-STAT3/Bcl-2 axis in ovarian cancer[J].Apoptosis,2017,22(4):544-557.