吉林大学学报(医学版) ›› 2022, Vol. 48 ›› Issue (6): 1474-1480.doi: 10.13481/j.1671-587X.20220613

• 基础研究 • 上一篇    下一篇

灵芝乙醇提取物对宫颈癌细胞生物学行为和JAK1/STAT3信号通路的影响

任璐(),曹芹雪,杨少琴   

  1. 河南大学淮河医院妇产科,河南 开封 475000
  • 收稿日期:2022-02-23 出版日期:2022-11-28 发布日期:2022-12-07
  • 通讯作者: 任璐 E-mail:hls_zhk@163.com
  • 作者简介:任 璐(1981-),女,河南省开封市人,副主任医师,主要从事妇科肿瘤基础和临床方面的研究。(E-mail:hls_zhk@163.com
  • 基金资助:
    河南省科技厅科研项目(182102311175)

Effects of ganoderma lucidum ethanol extract on biological behavior and JAK1/STAT3 signaling pathway of cervical cancer cells

Lu REN(),Qinxue CAO,Shaoqin YANG   

  1. Department of Obstetrics and Gynecology,Huaihe Hospital,Henan University,Kaifeng 475000,China
  • Received:2022-02-23 Online:2022-11-28 Published:2022-12-07
  • Contact: Lu REN E-mail:hls_zhk@163.com

摘要:

目的 探讨灵芝乙醇提取物(GLEE)对宫颈癌细胞生物学行为和Janus激酶1(JAK1)/信号传导和转录激活因子3(STAT3)信号通路的影响,并阐明其可能的作用机制。 方法 将人宫颈癌HeLa细胞随机分为对照组(给予完全培养液)和低、中及高剂量GLEE组(给予25、50、100 mg·L-1 GLEE)。MTT法检测各组细胞增殖抑制率,平板克隆实验检测各组克隆形成率,Transwell小室实验检测各组迁移细胞数和侵袭细胞数,实时荧光定量PCR(RT-qPCR)法检测各组细胞中JAK1和STAT3 mRNA表达水平,Western blotting法检测各组细胞中JAK1、磷酸化JAK1(p-JAK1)、STAT3、磷酸化STAT3(p-STAT3)、细胞因子信号传导抑制蛋白3(SOCS3)和活化STAT蛋白抑制因子1(PIAS1)蛋白表达水平。 结果 与对照组比较,低、中和高剂量GLEE组宫颈癌HeLa细胞增殖抑制率升高(P<0.05),细胞克隆形成率、迁移细胞数和侵袭细胞数降低(P<0.05),细胞中JAK1和STAT3 mRNA表达水平降低(P<0.05),细胞中p-JAK1和p-STAT3蛋白表达水平降低(P<0.05),细胞中SOCS3和PIAS1蛋白表达水平升高(P<0.05)。与低剂量GLEE组比较,中和高剂量GLEE组宫颈癌HeLa细胞增殖抑制率升高(P<0.05),细胞克隆形成率、迁移细胞数和侵袭细胞数降低(P<0.05),细胞中JAK1和STAT3 mRNA表达水平降低(P<0.05),细胞中p-JAK1和p-STAT3蛋白表达水平降低(P<0.05),细胞中SOCS3和PIAS1蛋白表达水平升高(P<0.05)。与中剂量GLEE组比较,高剂量GLEE组宫颈癌HeLa细胞增殖抑制率升高(P<0.05),细胞克隆形成率、迁移细胞数和侵袭细胞数降低(P<0.05),细胞中JAK1和STAT3 mRNA表达水平降低(P<0.05),细胞中p-JAK1和p-STAT3蛋白表达水平降低(P<0.05),细胞中SOCS3和PIAS1蛋白表达水平升高(P<0.05)。 结论 GLEE对宫颈癌HeLa细胞增殖、克隆形成、迁移和侵袭均有一定的抑制作用,其可能是通过上调细胞中SOCS3及PIAS1表达水平,进而影响JAK1/STAT3信号通路发挥作用。

关键词: 灵芝乙醇提取物, 宫颈肿瘤, Janus激酶1, 信号传导和转录激活因子3, 细胞因子信号传导抑制蛋白3

Abstract:

Objective To investigate the effects of ganoderma lucidum ethanol extract (GLEE) on the biological behavior and Janus kinase 1(JAK1)/signal transduction and transcription activator 3 (STAT3) signaling pathway of the cervical cancer cells, and to clarify its possible mechanisms. Methods The human cervical cancer HeLa cells were randomly divided into control group (given complete culture medium) and low, medium and high doses of GLEE groups(given 25, 50, and 100 mg·L-1 GLEE). The inhibitory rates of proliferation of the human cervical cancer HeLa cells in various groups were detected by MTT assay; the clone formation rates of the human cervical cancer HeLa cells in various groups were detected by plate cloning experiment; Transwell chamber assay was used to detect the numbers of migration cells and invasion cells in various groups; the expression levels of JAK1 and STAT3 mRNA in the human cervical cancer HeLa cells in various groups were detected by real-time fluorescence quantitative PCR (RT-qPCR) method; the expression levels of JAK1, phophorylated JAK1(p-JAK1),STAT3, phophorylated STAT3(p-STAT3), suppressor of cytokine signaling 3(SOCS3), and protein inhibitor of activated STAT1 (PIAS1) proteins in the human cervical cancer HeLa cells in various groups were detected by Western blotting method. Results Compared with control group, the inhibitory rates of proliferation of the cervical cancer HeLa cells in low, medium and high doses of GLEE groups were increased (P<0.05),the clone formation rates the cervical cancer HeLa cells were decreased (P<0.05),the numbers of migration cells were decreased (P<0.05), the numbers of invasion cells were decreased (P<0.05),the expression levels of JAK1 and STAT3 mRNA in the cervical cancer HeLa cells were decreased (P<0.05), and the expression levels of SOCS3 and PIAS1 proteins in the cervical cancer HeLa cells were increased(P<0.05). Compared with low dose of GLEE group, the inhibitory rates of proliferation of the cervical cancer HeLa cells in medium and high doses of GLEE groups were increased (P<0.05), the clone formation rates of the cervical cancer HeLa cells were decreased (P<0.05), the numbers of migration cells were decreased (P<0.05),the numbers of invasion cells were decreased (P<0.05), the expression levels of JAK1 and STAT3 mRNA in the cervical cancer HeLa cells were decreased (P<0.05),and the expression levels of SOCS3 and PIAS1 proteins in the cervical cancer HeLa cells were increased (P<0.05). Compared with midium dose of GLEE group, the inhibitory rate of proliferation of the cervical cancer HeLa cells in high dose of GLEE group was increased (P<0.05),the clone formation rate of the cervical cancer HeLa cells was decreased (P<0.05),the number of migration cells was decreased (P<0.05),the number of invasive cells was decreased (P<0.05), the expression levels of JAK1 and STAT3 mRNA in the cervical cancer HeLa cells were decreased (P<0.05),and the expression levels of SOCS3 and PIAS1 proteins in the cervical cancer HeLa cells were increased (P<0.05). Conclusion GLEE can inhibit the proliferation, cloning, migration and invasion of the cervical cancer HeLa cells to some extent, which may play a role by up-regulating the expression levels of SOCS3 and PIAS1 in the cells, thereby affecting the JAK1/STAT3 signaling pathway.

Key words: Ganoderma lucidum ethanol extract, Cervical neoplasms, Janus kinase 1, Signal transduction and transcription activator 3, Cytokine signaling inhibitor protein 3

中图分类号: 

  • R730.52