细胞外泛素对肝癌细胞增殖、侵袭和迁移能力的影响

张杨; 陈淑英; 徐光如

doi:10.3969/j.issn.1001-5256.2020.06.020

细胞外泛素对肝癌细胞增殖、侵袭和迁移能力的影响

DOI: 10.3969/j.issn.1001-5256.2020.06.020

1. 上海市浦东新区人民医院肿瘤科
2. 复旦大学附属华山医院检验科

基金项目:

上海市青年科技英才扬帆计划(19YF1405500)；

详细信息

中图分类号: R735.7
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出版历程
- 出版日期: 2020-06-20

Effect of extracellular ubiquitin on the proliferation,invasion,and migration of hepatoma cells

Research funding:

摘要

摘要:
目的通过体外细胞实验,探讨细胞外泛素对肝细胞癌细胞增殖、侵袭和迁移能力的影响。方法利用不同浓度的细胞外泛素（200、400、800 ng/ml）,分别干预肝癌细胞（HepG2）,采用CCK8法检测细胞的增殖情况;利用Transwell法检测不同浓度泛素对肝癌细胞侵袭能力的影响;利用划痕实验和蛋白免疫印迹检测细胞外泛素对肝癌细胞迁移能力的作用。计量资料多组间比较采用单因素方差分析,进一步两两比较采用LSD-t检验。结果 CCK8法检测结果显示,细胞外泛素干预后能明显促进肝癌细胞的增殖能力,并具有明显的浓度依赖性,且泛素干预浓度在400 ng/ml时作用最显著,在48、72、96 h相对吸光度值均明显高于对照组（P值均<0. 05）。Transwell实验结果显示,与对照组相比,不同浓度的细胞外泛素均可明显促进肝癌细胞的侵袭,且在400 ng/ml泛素干预组最显著[（134. 00±8. 18）个vs （347. 33±18. 90）个,t=17. 94,P <0. 001]。划痕实验和蛋白免疫印迹结果显示,400 ng/ml泛素能显著增加HepG2肝癌细胞的迁移能力。结论细胞...
- 癌,肝细胞 /
- 泛素 /
- 细胞增殖 /
- 肿瘤侵润 /
- 细胞运动
Abstract:
Objective To investigate the effect of extracellular ubiquitin on the proliferation,invasion,and migration of hepatoma cells by in vitro cell experiments. Methods The hepatoma cells( HepG2) were treated with extracellular ubiquitin at different concentrations( 200,400,and 800 ng/ml); CCK-8 assay was used to observe cell proliferation,Transwell assay was used to observe the effect of extracellular ubiquitin at different concentrations on the invasion ability of hepatoma cells,and wound healing assay and Western blotting were used to observe the effect of extracellular ubiquitin on the migration ability of hepatoma cells. A one-way analysis of variance was used for comparison of continuous data between multiple groups,and the least significant difference t-test was used for further comparison between two groups.Results The CCK-8 assay showed that extracellular ubiquitin significantly promoted the proliferation of hepatoma cells in a concentration-dependent manner,with the most significant effect at the concentration of 400 ng/ml,and the intervention group had a significantly higher relative absorbance value than the control group at 48,72,and 96 hours( all P < 0. 05). The Transwell assay showed that different concentrations of extracellular ubiquitin significantly promoted the invasion of hepatoma cells,with the most significant effect in the group treated by400 ng/ml extracellular ubiquitin( 134. 00 ± 8. 18 vs 347. 33 ± 18. 90,t = 17. 94,P < 0. 001). The Wound healing assay and Western blotting showed that 400 ng/ml extracellular ubiquitin significantly increased the migration ability of HepG2 cells. Conclusion Extracellular ubiquitin can significantly promote the proliferation,invasion,and migration of HepG2 cells in vitro in a concentration-dependent manner.
- carcinoma,hepatocellular /
- ubiquitin /
- cell proliferation /
- neoplasm invasiveness /
- cell movement

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参考文献(22)

[1] SHI Y,ZHAI B. A recent advance in image-guided locoregional therapy for hepatocellular carcinoma[J]. Gastrointest Tumors,2016,3(2):90-102.

[2] BALOGH J,VICTOR D 3rd,ASHAM EH,et al. Hepatocellular carcinoma:A review[J]. J Hepatocell Carcinoma,2016,3:41-53.

[3] WANG CC,IYER SG,LOW JK,et al. Perioperative factors affecting long-term outcomes of 473 consecutive patients undergoing hepatectomy for hepatocellular carcinoma[J]. Ann Surg Oncol,2009,16(7):1832-1842.

[4] LIU L,WANG Z,JIANG S,et al. Perioperative allogenenic blood transfusion is associated with worse clinical outcomes for hepatocellular carcinoma:A meta-analysis[J]. PLo S One,2013,8(5):e64261.

[5] SHIBA H,ISHIDA Y,WAKIYAMA S,et al. Negative impact of blood transfusion on recurrence and prognosis of hepatocellular carcinoma after hepatic resection[J]. J Gastrointest Surg,2009,13(9):1636-1642.

[6] XUN Y,TIAN H,HU L,et al. The impact of perioperative allogeneic blood transfusion on prognosis of hepatocellular carcinoma after radical hepatectomy:A systematic review and meta-analysis of cohort studies[J]. Medicine(Baltimore),2018,97(43):e12911.

[7] PATEL MB,PROCTOR KG,MAJETSCHAK M. Extracellular ubiquitin increases in packed red blood cell units during storage[J]. J Surg Res,2006,135(2):226-232.

[8] YAN L,CAI Q,XU Y. The ubiquitin-CXCR4 axis plays an important role in acute lung infection-enhanced lung tumor metastasis[J]. Clin Cancer Res,2013,19(17):4706-4716.

[9] ZHANG J,CHEN S,YAN Y,et al. Extracellular ubiquitin is the causal link between stored blood transfusion therapy and tumor progression in a melanoma mouse model[J]. J Cancer,2019,10(12):2822-2835.

[10] GILSON CR,KRAUS TS,HOD EA,et al. A novel mouse model of red blood cell storage and posttransfusion in vivo survival[J]. Transfusion,2009,49(8):1546-1553.

[11] ZHANG W,ZHU XD,SUN HC,et al. Depletion of tumor-associated macrophages enhances the effect of sorafenib in metastatic liver cancer models by antimetastatic and antiangiogenic effects[J]. Clin Cancer Res,2010,16(13):3420-3430.

[12] KOCH CG,LI L,SESSLER DI,et al. Duration of red-cell storage and complications after cardiac surgery[J]. N Engl J Med,2008,358(12):1229-1239.

[13] CATA JP,WANG H,GOTTUMUKKALA V,et al. Inflammatory response,immunosuppression,and cancer recurrence after perioperative blood transfusions[J]. Br J Anaesth,2013,110(5):690-701.

[14] ATAIL S,ARAD M,GLASNER A,et al. Blood transfusion promotes cancer progression:A critical role for aged erythrocytes[J]. Anesthesiology,2008,109(6):989-997.

[15] HAAS AL,WILKINSON KD. The large scale purification of ubiquitin from human erythrocytes[J]. Prep Biochem,1985,15(1-2):49-60.

[16] ZHU X,YU B,YOU P,et al. Ubiquitin released in the plasma of whole blood during storage promotes mRNA expression of Th2 cytokines and Th2-inducing transcription factors[J].Transfus Apher Sci,2012,47(3):305-311.

[17] SAINI V,MARCHESE A,MAJETSCHAK M. CXC chemokine receptor 4 is a cell surface receptor for extracellular ubiquitin[J]. J Biol Chem,2010,285(20):15566-15576.

[18] STEAGALL RJ,DANIELS CR,DALAL S,et al. Extracellular ubiquitin increases expression of angiogenic molecules and stimulates angiogenesis in cardiac microvascular endothelial cells[J]. Microcirculation,2014,21(4):324-332.

[19] SCOFIELD SL,AMIN P,SINGH M,et al. Extracellular ubiquitin:Role in myocyte apoptosis and myocardial remodeling[J].Compr Physiol,2015,6(1):527-560.

[20] MAJETSCHAK M,COHN SM,OBERTACKE U,et al. Therapeutic potential of exogenous ubiquitin during resuscitation from severe trauma[J]. J Trauma,2004,56(5):991-999.

[21] EARLE SA,PROCTOR KG,PATEL MB,et al. Ubiquitin reduces fluid shifts after traumatic brain injury[J]. Surgery,2005,138(3):431-438.

[22] MAJETSCHAK M,COHN SM,NELSON JA,et al. Effects of exogenous ubiquitin in lethal endotoxemia[J]. Surgery,2004,135(5):536-543.

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