[Multidrug resistant effect of alternative splicing form of MAD2 gene-MAD2beta on human gastric cancer cell].
OBJECTIVE
To 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.


METHODS
RNA 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.


RESULTS
A 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.


CONCLUSIONS
MAD2beta could increase the multidrug resistance of SGC7901 cell line.
Yin
Fang
F
Department of Gastroenterology, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China.
Hu
Wen-hua
WH
Qiao
Tai-dong
TD
Fan
Dai-ming
DM
D000230:Adenocarcinoma / Q000378:metabolism / Q000473:pathology*; D017398:Alternative Splicing; D000903:Antibiotics, Antineoplastic / Q000494:pharmacology; D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology; D002135:Calcium-Binding Proteins / Q000096:biosynthesis / Q000235:genetics*; D018797:Cell Cycle Proteins; D045744:Cell Line, Tumor; D004268:DNA-Binding Proteins / Q000096:biosynthesis / Q000235:genetics*; D004317:Doxorubicin / Q000494:pharmacology; D018432:Drug Resistance, Multiple; D019008:Drug Resistance, Neoplasm / Q000235:genetics; D006801:Humans; D064246:Mad2 Proteins; D016685:Mitomycin / Q000494:pharmacology; D012097:Repressor Proteins; D051899:Smad2 Protein; D013274:Stomach Neoplasms / Q000378:metabolism / Q000473:pathology*; D015534:Trans-Activators / Q000096:biosynthesis / Q000235:genetics*; D014162:Transfection; D014750:Vincristine / Q000494:pharmacology
D000903:Antibiotics, Antineoplastic; D000972:Antineoplastic Agents, Phytogenic; D002135:Calcium-Binding Proteins; D018797:Cell Cycle Proteins; D004268:DNA-Binding Proteins; C577697:MAD2L1 protein, human; D064246:Mad2 Proteins; D012097:Repressor Proteins; C497782:SMAD2 protein, human; D051899:Smad2 Protein; D015534:Trans-Activators; D016685:Mitomycin; D014750:Vincristine; D004317:Doxorubicin