医学分子生物学杂志2019，Vol.16Issue(5)：453-459,7.DOI:10.3870/j.issn.1672-8009.2019.05.014
番泻苷A抑制JAK2/STAT3通路的激活诱导前列腺癌细胞LNCap自噬性死亡
Sennoside A Inhibits AK2/STAT3 Pathway Activation to Induce Auto-phagic Death of Prostate Cancer Cell Line LNCap
摘要
Abstract
Objective To explore the effects of sennoside A ( SA) on autophagic death of prostate cancer cell line LNCap and its role mechanism. Methods Prostate cancer LNCap cells were cultured in vitro and treated with different concentrations of SA (5, 10, 20 μmol/L) . Cell survival rate and proliferation were detected by CCK-8 method. Apoptosis rate was detected by FCM method. LC3 distribution in cells was detected by immunofluorescence. Western blotting was used to detect the levels of apoptosis-related proteins, autophagy proteins and signaling transduction path-way-related proteins of Janus kinase 2-signal transducer and activator of transcription 3 ( JAK2/STAT3) . In vivo, nude mice models of transplanted tumor were established by subcutaneous injec-tion of prostate cancer cell LNCap, and intraperitoneally administered with different doses of SA (5, 10, 20 mg/kg/day), and they were continuously administered for 3 weeks. The tumor mass was measured after administration, and the levels of proliferating cell nuclear antigen ( Ki67 anti-gen, Ki67) and cysteine proteinase ( Caspase-3 ) were detected by immunohistochemistry, and the levels of autophagy-related proteins, JAK2 and STAT3 were detected by Western blot-ting. Results Compared with Ctrl, with the increase of SA concentration, the survival rate and proliferation rate of LNCap cells were decreased while the apoptosis rate of LNCap cells was in-creased, and the levels of Ki67 and proliferating cell nuclear antigen ( PCNA) were decreased while the levels of Caspase-3 and Caspase-9 were increased. Immunofluorescence results showed that compared with Ctrl, the distribution of LC3 in LNCap cells was increased with the increase of SA concentration. The results of in vivo experiments in nude mice showed that compared with Ctrl, with the increase of SA concentration, the tumor mass was decreased gradually, and the level of Ki67 was decreased while the level of caspase-3 was increased. The results of Western blotting in vitro and in vivo showed that compared with Ctrl, the levels of autophagy-related gene (Beclin1) and mi-crotubule-associated protein 1 light chain 3-II/microtubule-associated protein 1 light chain 3-I (LC3-II/LC3-I) were increased with the increase of SA concentration while the levels of p62, P-JAK2, P-STAT3 and c-Myc were decreased with the increase of SA concentration. Conclusion SA can promote the autophagic death of prostate cancer cell line LNCap by inhibiting JAK2/STAT3 sig-naling transduction and up-regulating levels of autophagy and apoptotic proteins.关键词
前列腺癌细胞/番泻苷A/JAK2/STAT3信号通路/自噬Key words
prostate cancer cell line/Sennoside A/JAK2/STAT3 signaling pathway/Auto-phagy分类
医药卫生引用本文复制引用
吴文婧,申涛,赵华,孙波..番泻苷A抑制JAK2/STAT3通路的激活诱导前列腺癌细胞LNCap自噬性死亡[J].医学分子生物学杂志,2019,16(5):453-459,7.基金项目
陕西省自然科学基金 (No. 2018JM7045) This work was supported by a grant from the Natural Science Foundation of Shaanxi Province (No. 2018JM7045) （No. 2018JM7045）
