Chinese Journal of Gastrointestinal Surgery 2012;15(4):388-391

Screening and identification of genes associated with multi-drug resistance in colonic cancer.

Jian-fang LI 1 ; Zhong ZHENG ; Bei-qin YU ; Ying QU ; Zheng-gang ZHU ; Bing-ya LIU

Affiliations

+expand

Country

China

Language

Chinese

Abstract

OBJECTIVETo identify novel multi-drug resistance-related genes, and to explore the mechanisms of multi-drug resistance.

METHODSMulti-drug resistant cell line Lovo/5-FU was established by incubation with increasing dose of 5-FU. The sensitivity to 5-FU and cis-diaminodichloroplatinum (CDDP) was measured by MTT assay. Two dimensional electrophoresis plus mass spectrum(2-DE/MS) was used to identify the differentially expressed protein between Lovo and Lovo/5-FU. The identified protein was then verified by Western blot analysis.

RESULTSThe IC50 concentrations of Lovo/5-FU to 5-FU and CDDP were increased by 31 and 3 times, compared with Lovo (both P<0.01). 2DE-MS showed that CAP-G and RhoGDI2 were up-regulated, whereas 6-PGL, DCI, Prdx-6 and Maspin were down-regulated in Lovo/5-FU. Western blot analysis confirmed that the expression levels of RhoGDI2 and CAP-G in Lovo/5-FU were increased by 6.14 and 2.98 fold respectively (both P<0.01), whereas Maspin was decreased to 5.2% of Lovo(P<0.01).

CONCLUSIONSMulti-gene and multi-pathway are involved in the development of multi-drug resistance of colorectal cancer cells. CAP-G, RhoGDI2 and Maspin are potential multi-drug resistant genes.