下调IL35通过上调自噬相关蛋白表达而抑制肝细胞癌侵袭和迁移

易言; 陈青松; 吴忠均

doi:10.16016/j.2097-0927.202102042

您当前的位置：

首页 >

文章列表页 >

下调IL35通过上调自噬相关蛋白表达而抑制肝细胞癌侵袭和迁移

基础医学 | 更新时间：2026-01-09

- 下调IL35通过上调自噬相关蛋白表达而抑制肝细胞癌侵袭和迁移
- Knockdown of IL35 inhibits migration and invasion of hepatocellular carcinoma by up-regulating autophagy-related proteins in HCC cells
- 陆军军医大学学报 2023年45卷第5期页码：441-449
- 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：
  
  [吴忠均，E-mail: wzjtcy@126.comWU Zhongjun, E-mail: wzjtcy@126.com
- 基金信息：
  
  重庆市留学人员创新支持计划(CX2017105);重庆市留学人员创新支持计划(CX2018146)
- DOI：10.16016/j.2097-0927.202102042
  中图分类号： R392.2;R73-37;R735.7
- 收稿：2021-02-06，
  
  修回：2023-01-17，
  
  纸质出版：2023-03-15
- 稿件说明：
移动端阅览
易言, 陈青松, 吴忠均. 下调IL35通过上调自噬相关蛋白表达而抑制肝细胞癌侵袭和迁移[J]. 陆军军医大学学报, 2023,45(5):441-449.

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.
易言, 陈青松, 吴忠均. 下调IL35通过上调自噬相关蛋白表达而抑制肝细胞癌侵袭和迁移[J]. 陆军军医大学学报, 2023,45(5):441-449. DOI： 10.16016/j.2097-0927.202102042.

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

SIEGEL R L, MILLER K D, JEMAL A. Cancer statistics, 2015[J]. CA A Cancer J Clin, 2015, 65(1): 5-29. DOI: 10.3322/caac.21254.

CHEN W Q, ZHENG R S, BAADE P D, et al . Cancer statistics in China, 2015[J ] . CA A Cancer J Clin, 2016, 66(2): 115-132. DOI: 10.3322/caac.21338.

GRANDHI M S, KIM A K, RONNEKLEIV-KELLY S M, et al . Hepatocellular carcinoma: from diagnosis to treatment[J ] . Surg Oncol, 2016, 25(2): 74-85. DOI: 10.1016/j.suronc.2016.03.002.

SAWANT D V, HAMILTON K, VIGNALI D A A. Interleukin-35: expanding its job profile[J]. J Interferon Cytokine Res, 2015, 35(7): 499-512. DOI: 10.1089/jir.2015.0015.

FU Y P, YI Y, CAI X Y, et al . Overexpression of interleukin-35 associates with hepatocellular carcinoma aggressiveness and recurrence after curative resection[J ] . Br J Cancer, 2016, 114(7): 767-776. DOI: 10.1038/bjc.2016.47.

LONG J, GUO H Y, CUI S C, et al . IL-35 expression in hepatocellular carcinoma cells is associated with tumor progression[J ] . Oncotarget, 2016, 7(29): 45678-45686. DOI: 10.18632/oncotarget.10141.

LIANG Y F, CHEN Q Q, DU W J, et al . Epstein-Barr virus-induced gene 3 (EBI3) blocking leads to induce antitumor cytotoxic T lymphocyte response and suppress tumor growth in colorectal cancer by bidirectional reciprocal-regulation STAT3 signaling pathway[J ] . Mediators Inflamm, 2016, 2016: 3214105. DOI: 10.1155/2016/3214105.

HUANG C B, LI N, LI Z X, et al . Tumour-derived interleukin 35 promotes pancreatic ductal adenocarcinoma cell extravasation and metastasis by inducing ICAM1 expression[J ] . Nat Commun, 2017, 8: 14035. DOI: 10.1038/ncomms14035.

TAO Q S, PAN Y, WANG Y P, et al . Regulatory T cells-derived IL-35 promotes the growth of adult acute myeloid leukemia blasts[J ] . Int J Cancer, 2015, 137(10): 2384-2393. DOI: 10.1002/ijc.29563.

LEVINE B, KROEMER G. Biological functions of autophagy genes: a disease perspective[J]. Cell, 2019, 176(1/2): 11-42. DOI: 10.1016/j.cell.2018.09.048.

LUO M J, LIU Y F, LIU N N, et al . Proscillaridin A inhibits hepatocellular carcinoma progression through inducing mitochondrial damage and autophagy[J ] . Acta Biochim Biophys Sin (Shanghai), 2021, 53(1): 19-28. DOI: 10.1093/abbs/gmaa139.

TAKAMURA A, KOMATSU M, HARA T, et al . Autophagy-deficient mice develop multiple liver tumors[J ] . Genes Dev, 2011, 25(8): 795-800. DOI: 10.1101/gad.2016211.

OSMAN N A A, ABD EL-REHIM D M, KAMAL I M. Defective Beclin-1 and elevated hypoxia-inducible factor (HIF)-1α expression are closely linked to tumorigenesis, differentiation, and progression of hepatocellular carcinoma[J]. Tumor Biol, 2015, 36(6): 4293-4299. DOI: 10.1007/s13277-015-3068-0.

LI L, LIU J D, GAO G D, et al . Puerarin 6″-O-xyloside suppressed HCC via regulating proliferation, stemness, and apoptosis with inhibited PI3K/AKT/mTOR[J ] . Cancer Med, 2020, 9(17): 6399-6410. DOI: 10.1002/cam4.3285.

WANG R X, YU C R, DAMBUZA I M, et al . Interleukin-35 induces regulatory B cells that suppress autoimmune disease[J ] . Nat Med, 2014, 20(6): 633-641. DOI: 10.1038/nm.3554.

HAO S N, CHEN X, WANG F, et al . Breast cancer cell-derived IL-35 promotes tumor progression via induction of IL-35-producing induced regulatory T cells[J ] . Carcinogenesis, 2018, 39(12): 1488-1496. DOI: 10.1093/carcin/bgy136.

WU W, JIANG H, LI Y, et al . IL-35 expression is increased in laryngeal squamous cell carcinoma and in the peripheral blood of patients[J ] . Oncol Lett, 2017, 13(5): 3303-3308. DOI: 10.3892/ol.2017.5858.

康海岩, 杨庭显, 张磊. 甲状腺癌患者Gal-3、IL-17、IL-35、anti-TgAb的变化及期意义[J]. 西北国防医学杂志, 2017, 38(8): 505-508.

KANG H Y, YANG T X, ZHANG L. Significance of serum Gal-3, IL-17, IL-35 and anti-TgAb in patients with thyroid carcinoma[J]. Med J Natl Defend Forces Northwest China, 2017, 38(8): 505-508.

刘若时, 李慧, 吴继周, 等. IL-35、IL-22与广西肝癌家族聚集性的相关性研究[J]. 广西医科大学学报, 2018, 35(2): 170-173.

LIU R S, LI H, WU J Z, et al. The relationship between IL-35, IL-22 and familial aggregation of hepatocellular carcinoma in Guangxi[J ] . J Guangxi Med Univ, 2018, 35(2): 170-173.

ROSSITER H, KÖNIG U, BARRESI C, et al . Epidermal keratinocytes form a functional skin barrier in the absence of Atg7 dependent autophagy[J ] . J Dermatol Sci, 2013, 71(1): 67-75. DOI: 10.1016/j.jdermsci.2013.04.015.

ZHANG X G, JIN L, TIAN Z, et al . Nitric oxide inhibits autophagy and promotes apoptosis in hepatocellular carcinoma[J ] . Cancer Sci, 2019, 110(3): 1054-1063. DOI: 10.1111/cas.13945.

JI Y, LI L, MA Y X, et al . Quercetin inhibits growth of hepatocellular carcinoma by apoptosis induction in part via autophagy stimulation in mice[J ] . J Nutr Biochem, 2019, 69: 108-119. DOI: 10.1016/j.jnutbio.2019.03.018.

LI T T, ZHU D, MOU T, et al . IL-37 induces autophagy in hepatocellular carcinoma cells by inhibiting the PI3K/AKT/mTOR pathway[J ] . Mol Immunol, 2017, 87: 132-140. DOI: 10.1016/j.molimm .2017.04.010.

NIETO M A, HUANG R Y J, JACKSON R A, et al . Emt: 2016[J ] . Cell, 2016, 166(1): 21-45. DOI: 10.1016/j.cell.2016.06.028.

DONG Y Y, WU Z F, HE M Y, et al . ADAM9 mediates the interleukin-6-induced Epithelial-Mesenchymal transition and metastasis through ROS production in hepatoma cells[J ] . Cancer Lett, 2018, 421: 1-14. DOI: 10.1016/j.canlet.2018.02.010.

浏览量

下载量

CSCD

文章被引用时，请邮件提醒。

提交

工具集

关联资源

IFIT5在肝细胞癌中表达上调并促进肝癌细胞侵袭和迁移

乙醇通过促进上皮间质转化增强结直肠癌细胞的侵袭转移能力

沉默NUPR1降低人多发性骨髓瘤U266细胞自噬并促进其迁移和侵袭

CTP-NPRL2融合蛋白对肾癌原代细胞迁移侵袭能力的抑制及机制研究

IBP通过上皮间质转化促进结肠癌细胞迁移和侵袭