Protein kinase Cβ Ⅱ promotes invasion of hepatocellular carcinoma by modulating autocrine of IL-6

刘　敏; 陈添亮; 郭　娟; 高洪泉; 巫佳翠; 王玉孝

doi:10.19401/j.cnki.1007-3639.2021.04.005

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Protein kinase Cβ Ⅱ promotes invasion of hepatocellular carcinoma by modulating autocrine of IL-6

更新时间：2026-01-27

- Protein kinase Cβ Ⅱ promotes invasion of hepatocellular carcinoma by modulating autocrine of IL-6
- China Oncology Vol. 31, Issue 4, Pages: 268-276(2021)
- 作者机构：
  
  1. 厦门医学院基础医学部,福建,厦门,361023
  2. 机能与临床转化福建省高等学校重点实验室,福建,厦门,361023
  3. 厦门医学院临床医学系,福建,厦门,361023
  4. 厦门医学院附属第二医院病理科,福建,厦门,361023
- 作者简介：
- 基金信息：
- DOI：10.19401/j.cnki.1007-3639.2021.04.005
  CLC： R735.7
- Published Online：29 April 2021，
  
  Published：29 April 2021
- 稿件说明：
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刘　敏, 陈添亮, 郭　娟, 伊雪, 高洪泉, 巫佳翠, 王玉孝 . 蛋白激酶CβⅡ通过IL-6自分泌途径促进肝细胞癌侵袭的研究[J]. 中国癌症杂志, 2021, 31(4): 268-276.

刘　敏, 陈添亮, 郭　娟, et al. Protein kinase Cβ Ⅱ promotes invasion of hepatocellular carcinoma by modulating autocrine of IL-6[J]. China Oncology, 2021, 31(4): 268-276.
刘　敏, 陈添亮, 郭　娟, 伊雪, 高洪泉, 巫佳翠, 王玉孝 . 蛋白激酶CβⅡ通过IL-6自分泌途径促进肝细胞癌侵袭的研究[J]. 中国癌症杂志, 2021, 31(4): 268-276. DOI： 10.19401/j.cnki.1007-3639.2021.04.005.

刘　敏, 陈添亮, 郭　娟, et al. Protein kinase Cβ Ⅱ promotes invasion of hepatocellular carcinoma by modulating autocrine of IL-6[J]. China Oncology, 2021, 31(4): 268-276. DOI： 10.19401/j.cnki.1007-3639.2021.04.005.

摘要

背景与目的：肿瘤微环境中存在的炎性因子参与肿瘤的生长及转移，蛋白激酶CβⅡ（protein kinase CβⅡ，PKCβⅡ）能否通过调节炎性因子参与肝细胞癌（hepatocellular carcinoma，HCC）的侵袭尚不清楚。研究PKCβⅡ对HCC侵袭的作用及相关机制，深入探讨PKCβⅡ与肿瘤炎性微环境之间的关系。方法：收集PKCβⅡ高表达细胞的条件培养基处理HCC细胞，采用transwell法检测细胞的侵袭情况，采用细胞因子抗体芯片检测上清液中细胞因子的变化，采用酶联免疫吸附测定（enzyme-linked immunosorbent assay，ELISA）和实时荧光定量聚合酶链反应（real-time fluorescence quantitative polymerase chain reaction，RTFQ-PCR）检测PKCβⅡ高表达对白细胞介素-6（interleukin-6，IL-6）上清液中含量及转录的影响，采用transwell法检测IL-6中和抗体及IL-6受体（IL-6 receptor，IL-6R）中和抗体对PKCβⅡ高表达诱导HCC细胞侵袭的影响，采用Western blot检测信号转导与转录激活因子3（signal transducer and activator of transcription 3，STAT3）、P-STAT3的蛋白表达，采用非参数Spearman检验分析HCC组织中PKCβⅡ和IL-6表达的相关性。结果：采用PKCβⅡ高表达细胞的条件培养基处理可以提高HCC细胞Huh7（P0.01）和Hep3B（P0.05）的侵袭能力；PKCβⅡ高表达显著提高上清液中IL-6的含量（P0.01）及IL-6的转录（P0.01）；IL-6中和抗体及IL-6R中和抗体均可以抑制PKCβⅡ介导的HCC细胞侵袭能力的增加（P0.01），P-STAT3蛋白表达升高（P0.05）；HCC组织中PKCβⅡ和IL-6的表达呈正相关（r=0.697，P0.01）。结论：PKCβⅡ通过IL-6自分泌途径促进HCC细胞的侵袭。

Abstract

Background and purpose: Multiple proinflammatory cytokines in tumorigenic microenvironment are involved in the acquired capability for invasive growth and metastasis. It is not clear whether protein kinase CβⅡ(PKCβⅡ) can modulate the secretion of proinflammatory cytokines to promote the progression of hepatocellular carcinoma (HCC). This study aimed to explore the function and mechanism of PKCβⅡ in the invasion of HCC to investigate the relationship between PKCβⅡ and tumor inflammatory microenvironment. Methods: The effect of conditioned medium from PKCβⅡ-overexpressing cells on the invasion of HCC cells was detected by the transwell assay. Serum-free culture supernatants were analyzed by cytokine arrays. The effect of PKCβⅡ overexpression on the expression of interleukin-6 (IL-6) in supernatants and at mRNA level was examined by enzyme- linked immunosorbent assay (ELISA) and real-time fluorescence quantitative polymerase chain reaction (RTFQ-PCR) respectively. The effects of IL-6 neutralizing antibody and IL-6 receptor (IL-6R) neutralizing antibody on PKCβⅡ overexpression-induced invasion of HCC cells were detected by the transwell assay. The expressions of signal transducer and activator of transcription 3 (STAT3) and P-STAT3 were detected by Western blot. Correlation of PKCβⅡ with the expression of IL-6 in clinical HCC cases was analyzed using the nonparametric Spearman test. Results: Conditioned medium obtained from PKCβⅡ-overexpressing cells significantly induced the invasiveness of Huh7 (P0.01) and Hep3B (P0.05)

and the upregulated secretion of IL-6 (P0.01) and mRNA level (P0.01) upon PKCβⅡ overexpression were observed respectively. Administrations of IL-6 neutralizing antibody and IL-6R neutralizing antibody blocked PKCβⅡ overexpression-induced invasion of HCC cells (P0.01). The protein level of P-STAT3 increased significantly upon PKCβⅡ overexpression (P0.05). Immunohistochemical analysis also presented a strong positive correlation between PKCβⅡ and IL-6 expression in clinical HCC samples (r=0.697

P0.01). Conclusion: PKCβⅡ promotes the invasion of HCC by modulating autocrine of IL-6.

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