Knockdown of IL35 inhibits migration and invasion of hepatocellular carcinoma by up-regulating autophagy-related proteins in HCC cells

Yan YI; Qingsong CHEN; Zhongjun WU

doi:10.16016/j.2097-0927.202102042

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Knockdown of IL35 inhibits migration and invasion of hepatocellular carcinoma by up-regulating autophagy-related proteins in HCC cells

Basic Medicine | 更新时间：2026-01-09

- Knockdown of IL35 inhibits migration and invasion of hepatocellular carcinoma by up-regulating autophagy-related proteins in HCC cells
- Journal of Army Medical University Vol. 45, Issue 5, Pages: 441-449(2023)
- 作者机构：
  
  400016 重庆，重庆医科大学附属第一医院肝胆外科
- 作者简介：
- 基金信息：
  
  the Innovation Support Program for Overseas Students of Chongqing(CX2017105);the Innovation Support Program for Overseas Students of Chongqing(CX2018146)
- DOI：10.16016/j.2097-0927.202102042
  CLC： R392.2;R73-37;R735.7
- Received：06 February 2021，
  
  Revised：2023-01-17，
  
  Published：15 March 2023
- 稿件说明：
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Yan YI, Qingsong CHEN, Zhongjun WU. Knockdown of IL35 inhibits migration and invasion of hepatocellular carcinoma by up-regulating autophagy-related proteins in HCC cells[J]. Journal of Army Medical University, 2023, 45(5): 441-449.
DOI：

Yan YI, Qingsong CHEN, Zhongjun WU. Knockdown of IL35 inhibits migration and invasion of hepatocellular carcinoma by up-regulating autophagy-related proteins in HCC cells[J]. Journal of Army Medical University, 2023, 45(5): 441-449. DOI： 10.16016/j.2097-0927.202102042.

摘要

目的

探讨白细胞介素35(interleukin 35

IL35)在肝癌中的表达及其临床意义、潜在的分子机制。

方法

收集手术确诊的肝细胞肝癌和癌旁组织40对。采用RT-qPCR和Western blot检测IL35在肝癌和癌旁组织中的表达，并分析IL35与各临床指标的相关性。Western blot检测IL35在肝癌细胞中的表达。慢病毒转染下调IL35。CCK-8和集落形成检测细胞增殖，Transwell实验检测侵袭和迁移，Western blot检测细胞自噬和EMT相关指标。

结果

IL35高表达于肝癌组织中(

0.01)；与HL7702肝细胞相比，IL35蛋白表达在Hep3B、Huh7、SMMC7721、Bel7402细胞中升高，且Hep3B表达最高(

0.01); 高表达与血管侵犯相关(

=0.0033); Transwell实验表明下调L35能有效抑制肝癌细胞侵袭和转移(

0.01)；CCK-8和平板克隆实验表明，下调IL35能有效抑制肝癌细胞增殖(

0.01)；Western blot结果显示，下调IL35能促进LC3BII、Beclin1表达，抑制p62表达(

0.01)，同时促进E-cadherin表达并抑制N-cadherin、Vimentin的表达(

0.01)。

结论

IL35高表达于肝细胞肝癌组织和肝癌细胞中，其表达与肝癌的血管侵犯相关；下调IL35能通过诱导自噬达到抑制肝癌细胞的侵袭和迁移的目的。

Abstract

Objective

To investigate the expression

clinical significance and potential molecular mechanism of interleukin 35 (IL35) in hepatocellular carcinoma (HCC).

Methods

A total of 40 pairs of HCC and adjacent tissues were collected. RT-qPCR and Western blotting were used to detect the expression of IL35 in the tissues

and the correlation of IL35 with clinical indicators was analyzed. The expression of IL35 was detected in normal liver cell line LO2 and 4 strains of HCC Huh7

Hep3B

SMMC7721 and Bel7402 cells by Western blotting. Lentiviral infection was used to down-regulate the expression of IL35 in Hep3B cells. Then CCK-8 assay and colony formation were employed to detect cell proliferation

Transwell assay was adopted to detect cell invasion and migration

and Western blotting was performed to detect autophagy and epithelial-mesenchymal transition (EMT) related indicators.

Results

IL35 was highly expressed in HCC tissue (

0.01). Compared with HL7702 cells

IL35 was highly expressed in Hep3B

Huh7

SMMC7721 and BEL7402 cells

with the highest expression in Hep3B cells (

0.01). The high expression was associated with vascular invasion (P=0.0033). Transwell assay showed that down-regulation of IL35 effectively inhibited the invasion and metastasis of Hep3B cells (

0.01); CCK-8 assay and plate cloning assay indicated that down-regulation of IL35 effectively inhibited the proliferation of Hep3B cells (

0.01). Western blotting results suggested that down-regulation of IL35 promoted the expression of LC3BII and Beclin1

inhibited the expression of p62 (

0.01)

enhanced the expression of E-cadherin and suppressed the expression of N-cadherin and vimentin (

0.01).

Conclusion

IL35 is highly expressed in HCC tissues and cells

and its expression is related to vascular invasion of HCC. Down-regulation of IL35 can inhibit the invasion and migration of HCC cells by inducing autophagy.

关键词

Keywords

references

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