The mechanism of 5-Fu-based drug resistance in DNA mismatch repair deficient colorectal cancer HCT-116 cells

WANG Jing; FANG Hongliang; HUANG Jinlu; GUO Cheng

doi:10.3969/j.issn.1006-0111.2017.02.006

Message Board

Respected readers, authors and reviewers, you can add comments to this page on any questions about the contribution, review, editing and publication of this journal. We will give you an answer as soon as possible. Thank you for your support!

Name

E-mail

Phone

Title

Content

Verification Code

Objective To study the mechanisms of the drug resistance of DNA mismatch repair (MMR) deficient colorectal cancer (CRC) HCT-116 to 5-fluorouracil (5-Fu). Methods MLH1 deficiency HCT-116 cells were transfected with pcDNA3.1-MLH1 Vector.The expression of MLH1 was detected by Western blot.The change of resistance against 5-Fu was examined by detecting the cell viability with CCK-8 kits.The expression of CD133 (cancer stem cell marker) and CK8 & CK20 (cell differentiation marker) were detected by flow cytometry. Results Comparing to HCT-116 control group, the viability of HCT-116 cells was markedly decreased (P<0.01) after stable expressing MLH1, accompanied by the down-regulated expression of CD133 on the cell surface.Moreover, the up-regulation of cell differentiation marker CK8 and CK20 was observed in HCT-116 cells with stable expressing MLH1. Conclusion Our data indicated that the expression of MLH1 was associated with down-regulated CD133⁺ stem-like cells in colorectal cancer HCT-116 with MLH1 deficiency.Therefore, CD133⁺ stem-like cells may related to the drug resistance of MMR deficiency tumor.This study provides a possible theory to explain the 5-FU resistance in the colorectal cancer patients with MMR deficiency.

The mechanism of 5-Fu-based drug resistance in DNA mismatch repair deficient colorectal cancer HCT-116 cells

1. Sixth People's Hospital Affiliated to Shanghai JiaoTong University, Shanghai 200233, China

2. Department of Immunology, Faculty of Basic Medicine, Second Military Medical University, Shanghai 200433, China

Received Date: 2016-10-28

Rev Recd Date: 2017-01-09

Key words:

Abstract: Objective To study the mechanisms of the drug resistance of DNA mismatch repair (MMR) deficient colorectal cancer (CRC) HCT-116 to 5-fluorouracil (5-Fu). Methods MLH1 deficiency HCT-116 cells were transfected with pcDNA3.1-MLH1 Vector.The expression of MLH1 was detected by Western blot.The change of resistance against 5-Fu was examined by detecting the cell viability with CCK-8 kits.The expression of CD133 (cancer stem cell marker) and CK8 & CK20 (cell differentiation marker) were detected by flow cytometry. Results Comparing to HCT-116 control group, the viability of HCT-116 cells was markedly decreased (P<0.01) after stable expressing MLH1, accompanied by the down-regulated expression of CD133 on the cell surface.Moreover, the up-regulation of cell differentiation marker CK8 and CK20 was observed in HCT-116 cells with stable expressing MLH1. Conclusion Our data indicated that the expression of MLH1 was associated with down-regulated CD133⁺ stem-like cells in colorectal cancer HCT-116 with MLH1 deficiency.Therefore, CD133⁺ stem-like cells may related to the drug resistance of MMR deficiency tumor.This study provides a possible theory to explain the 5-FU resistance in the colorectal cancer patients with MMR deficiency.

HTML

Reference (17)

[1]	Brenner H,Kloor M,Pox CP.Colorectal cancer[J].Lancet,2014,383(9927):1490-1502.
[2]	Chen Q,Liu Z,Cheng L, et al.An analysis of incidence and mortality of colorectal cancer in China,2003~2007[J].China Cancer,2012,21(3):179-182.
[3]	Hsieh P,Yamane K.DNA mismatch repair:molecular mechanism,cancer,and ageing[J].Mech Ageing Dev,2008,129(7-8):391-407.
[4]	Hewish M,Lord CJ,Martin SA,et al.Mismatch repair deficient colorectal cancer in the era of personalized treatment[J].Nat Rev Clin Oncol,2010,7(4):197-208.
[5]	Sinicrope FA.DNA mismatch repair and adjuvant chemotherapy in sporadic colon cancer[J].Nat Rev Clin Oncol,2010,7(3):174-177.
[6]	Boland GM,Chang GJ,Haynes AB,et al.Association between adherence to National Comprehensive Cancer Network treatment guidelines and improved survival in patients with colon cancer[J].Cancer,2013,119(8):1593-1601.
[7]	祝利民,沈克平,周浩,等.胃肠安及四藤方对人结肠癌细胞株干细胞CD133⁺的影响[J].上海交通大学学报(医学版),2016,36(2):161-165.
[8]	Taverna P,Liu L,Hanson AJ,et al.Characterization of MLH1 and MSH2 DNA mismatch repair proteins in cell lines of the NCI anticancer drug screen[J].Cancer Chemother Pharmacol,2000,46(6):507-516.
[9]	Rosen JM,Jordan CT.The increasing complexity of the cancer stem cell paradigm[J].Science,2009,324(5935):1670-1673.
[10]	Zeki SS,Graham TA,Wright NA.Stem cells and their implications for colorectal cancer[J].Nat Rev Gastroenterol Hepatol,2011,8(2):90-100.
[11]	O'Brien CA,Pollett A,Gallinger S,et al.A human colon cancer cell capable of initiating tumour growth in immunodeficient mice[J].Nature,2007,445(7123):106-110.
[12]	Ricci-Vitiani L,Lombardi DG,Pilozzi E,et al.Identification and expansion of human colon-cancer-initiating cells[J].Nature,2006,445(7123):111-115.
[13]	Dallas NA,Xia L,Fan F, et al.Chemoresistant colorectal cancer cells,the cancer stem cell phenotype,and increased sensitivity to insulin-like growth factor-I receptor inhibition[J].Cancer Res,2009,69(5):1951-1957.
[14]	Ma S,Lee TK,Zheng BJ,et al.CD133⁺ HCC cancer stem cells confer chemoresistance by preferential expression of the Akt/PKB survival pathway[J].Oncogene,2008,27(12):1749-1758.
[15]	Yasuda H,Tanaka K,Saigusa S,et al.Elevated CD133,but not VEGF or EGFR,as a predictive marker of distant recurrence after preoperative chemoradiotherapy in rectal cancer[J].Oncol Rep,2009,22(4):709-717.
[16]	Sinicrope FA,Mahoney MR,Smyrk TC, et al.Prognostic impact of deficient DNA mismatch repair in patients with stage Ⅲ colon cancer from a randomized trial of FOLFOX-based adjuvant chemotherapy[J].J Clin Oncol,2013,31(29):3664- 3672.
[17]	Sargent DJ,Marsoni S,Monges G,et al.Defective mismatch repair as a predictive marker for lack of efficacy of fluorouracil-based adjuvant therapy in colon cancer[J].J Clin Oncol,2010,28(20):3219-3226.

Message Board

The mechanism of 5-Fu-based drug resistance in DNA mismatch repair deficient colorectal cancer HCT-116 cells

doi: 10.3969/j.issn.1006-0111.2017.02.006

Abstract

References

Proportional views

通讯作者: 陈斌, [email protected]

Article Metrics

Related

Proportional views