褪黑素通过抑制JAK2/STAT3通路抑制IL-6诱导的卵巢癌SKOV3细胞上皮间质转化

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摘要目的探究褪黑素对白细胞介素(IL)-6诱导的卵巢癌SKOV3细胞上皮间质转化(EMT)的作用和机制。方法首先用不同浓度的褪黑素(1、2、4、6和8 mM)与50 ng/mL重组IL-6蛋白处理SKOV3细胞,CCK-8检测细胞活力,选择两个褪黑素浓度,进行后续实验。然后将SKOV3细胞随机分成四组:对照组(不进行任何处理)、IL-6组(加入50 ng/mL IL-6)、IL-6+褪黑素低剂量组(同时加入50 ng/mL IL-6和2 mM褪黑素)、IL-6+褪黑素高剂量组(同时加入50 ng/mL IL-6和4 mM褪黑素)。IL-6加入48 h后,Transwell检测迁移细胞数目,Real-time PCR检测E-cadherin和Vimentin mRNA表达,Western blot检测E-cadherin、Vimentin、Janus激酶2(JAK2)、磷酸化JAK2(p-JAK2)、信号传导和转录激活因子3(STAT3)和磷酸化STAT3(p-STAT3)蛋白表达。结果褪黑素能够抑制IL-6增加的细胞活力,且呈现剂量依赖性。后续选择2 mM和4 mM褪黑素进行实验。与对照组相比,IL-6组中迁移细胞数目、Vimentin mRNA和蛋白水平、p-JAK2/JAK2和p-STAT3/STAT3增加,E-cadherin mRNA和蛋白水平降低,差异均有统计学意义(P均<0.05);与IL-6组相比,IL-6+褪黑素组迁移细胞数目、Vimentin mRNA和蛋白水平、p-JAK2/JAK2和p-STAT3/STAT3降低,E-cadherin mRNA和蛋白水平增加,且高剂量组中变化更加明显,差异均具有统计学意义(P均<0.05)。结论褪黑素能够抑制IL-6诱导的卵巢癌SKOV3细胞EMT,可能是通过抑制JAK2/STAT3通路发挥作用。

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	谢婷婷
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	韩碧洁
	于月成

关键词 ：褪黑素, IL-6, SKOV3细胞, 上皮间质转化, JAK2/STAT3

Abstract：Objective This study aimed to investigate the role and mechanism of melatonin on interleukin (IL)-6-induced epithelial-to-mesenchymal transition (EMT) in ovarian cancer SKOV3 cells. Methods SKOV3 cells were treated with different concentrations of melatonin (1, 2, 4, 6 and 8 mM) and 50 ng/mL IL-6. Two concentrations of melatonin were selected by cell viability determined using CCK-8 assay. Then, SKOV3 cells were randomly divided into 4 groups: the control group (treated with nothing), the IL-6 group (treated with 50 ng/mL IL-6), the IL-6+melatonin-low group (co-treated with 50 ng/mL IL-6 and 2 mM melatonin), and the IL-6+melatonin-high group (co-treated with 50 ng/mL IL-6 and 4 mM melatonin). After stimulation with IL-6 for 48 h, migrated cell number was detected by transwell. The mRNA expressions of E-cadherin and Vimentin were analyzed by real-time PCR. The protein levels of E-cadherin, Vimentin, Janus Kinase 2 (JAK2), phosphorylation of JAK2 (p-JAK2), Signal Transducer and Activator of Transcription 3 (STAT3), phosphorylation of STAT3 (p-STAT3) were measured by western blot. Results Melatonin could inhibit IL-6-elevated cell viability in a dose-dependent manner. 2 mM and 4 mM of melatonin were used in the following study. Compared with the control group, migrated cell numbers, the mRNA and protein levels of Vimentin, p-JAK2/JAK2 and p-STAT3/STAT3 were increased, and the mRNA and protein levels of E-cadherin were decreased in the IL-6 group (all P<0.05). Compared with the IL-6 group, migrated cell numbers, the mRNA and protein levels of Vimentin, p-JAK2/JAK2 and p-STAT3/STAT3 were reduced, and the mRNA and protein levels of E-cadherin were elevated in the IL-6+melatonin group, and these changes were more obvious in the IL-6+melatonin-high group (all P<0.05). Conclusion Melatonin can suppress IL-6-induced EMT in ovarian cancer SKOV3 cells, which may be related to the inactivation of JAK2/STAT3 signaling.

Key words： Melatonin IL-6 SKOV3 cells Epithelial-to-mesenchymal transition JAK2/STAT3

收稿日期: 2018-11-05

通讯作者: 于月成(yuecheng_yyc@163.com)

引用本文:

谢婷婷, 李雯惠, 苏瑛, 韩碧洁, 于月成. 褪黑素通过抑制JAK2/STAT3通路抑制IL-6诱导的卵巢癌SKOV3细胞上皮间质转化[J]. 中国生育健康杂志, 2019, 30(5): 431-435.
XIE Tingting, LI Wenhui, SU Ying, HAN Bijie, YU Yuecheng. Melatonin attenuates IL-6-induced epithelial-to-mesenchymal transition via inhibition of JAK2/STAT3 signaling pathway in ovarian cancer SKOV3 cells. Chinese Journal of Reproductive Health, 2019, 30(5): 431-435.

链接本文:

http://cjrh.bjmu.edu.cn/CN/ 或 http://cjrh.bjmu.edu.cn/CN/Y2019/V30/I5/431

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