Multidrug resistant effect of alternative splicing form of MAD2 gene-MAD2beta on human gastric cancer cell.
- Author:
Fang YIN
1
;
Wen-hua HU
;
Tai-dong QIAO
;
Dai-ming FAN
Author Information
- Publication Type:Journal Article
- MeSH: Adenocarcinoma; metabolism; pathology; Alternative Splicing; Antibiotics, Antineoplastic; pharmacology; Antineoplastic Agents, Phytogenic; pharmacology; Calcium-Binding Proteins; biosynthesis; genetics; Cell Cycle Proteins; Cell Line, Tumor; DNA-Binding Proteins; biosynthesis; genetics; Doxorubicin; pharmacology; Drug Resistance, Multiple; Drug Resistance, Neoplasm; genetics; Humans; Mad2 Proteins; Mitomycin; pharmacology; Repressor Proteins; Smad2 Protein; Stomach Neoplasms; metabolism; pathology; Trans-Activators; biosynthesis; genetics; Transfection; Vincristine; pharmacology
- From: Chinese Journal of Oncology 2004;26(4):201-204
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo study the effect of alternative splicing form -MAD2beta of mitotic arrest deficient protein 2 (MAD2) on the formation of multidrug resistance in human gastric adenocarcinoma cell SGC7901.
METHODSRNA was extracted from a multidrug resistance cell line SGC7901/ADR. The full-length MAD2beta cDNA was obtained by RT-PCR and cloned into the pUCm-T vector, and then recombined into the eukaryotic expression vector pcDNA3.1 in forward direction. Subsequently, pcDNA3.1/MAD2beta vectors were then transfected into SGC7901 cells by lipofectamine. Sensitivity to drug was detected by MTT assay. Cell cycle alteration and intracellular fluorescence intensity were determined by FACS.
RESULTSA fragment of 0.53 Kb was obtained and confirmed by DNA sequencing which was a new alternative splicing form of MAD2 named as MAD2beta. pcDNA3.1/MAD2beta transfected SGC7901 cells (SGC7901/MAD2beta) were more resistant to ADR, VCR and MMC than the control cells (SGC7901/pcDNA3.1), and also ADR fluorescence intensity of SGC7901/MAD2beta cells was lower (P < 0.05) than that of SGC7901/pcDNA3.1 cells.
CONCLUSIONMAD2beta could increase the multidrug resistance of SGC7901 cell line.
