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AIM:To investigate the expression level of ZNRD1 gene ingastric cancer cells SGC7901 and gastric cancer MDR(multidrug resistant) cells SGC7901/VCR,and to observethe drug sensitizing and proliferation effect of ZNRD1antisense nucleic acid transduction on SGC7901/VCR cells.METHODS:Amplification of sequences encoding ZNRD1from SGC7901/VCR cDNA by PCR.The levels of ZNRD1mRNA expression were demonstrated using semiquantitativereverse transcription polymerase chain reaction (RT-PCR).Eukaryotic expression vector pcDNA3.1-anti ZNRD1 wasconstructed and transfected into SGC7901/VCR cells bylipofectamine.Immunochemical method was used to detectthe expression of protein in SGC7901/VCR cells andtransfectants.The cell cycle alteration and the intracellularadriamycin (ADM) accumulation were observed by FACS.Growth curve and drug sensitization of cells for vincristine(VCR) were analyzed with MTT assay.RESULTS:We cloned the open reading frame of full-lengthZNRD1.The expression of ZNRD1 showed higher inSGC7901/VCR than in SGC7901 cells.The antisense ZNRD1drug-resistant clones were selected after gene transfection.Immunochemical results showed that the expression levelof ZNRD1 protein was lower in anti ZNRD1-SGC7901/VCRcells than that in non-transfectants.Comparing to SGC7901/VCR and pcDNA3.1-SGC7901/VCR,anti ZNRD1-SGC7901/VCR showed gradually accumulated in G_1 phase,with aconcomitant decrease of cell population in S phase.FACSalso suggested intracellular ADM accumulation increased2fold in SGC7901/VCR cells after transfected with antisenseZNRD1.MTT assay showed that transfectants cellsproliferation was lagged and more sensitive to VCR thannon-transfectants.CONCLUSION:ZNRD1 gene displayed highly expressionin VCR resistant gastric cancer cells.Expression of ZNRD1protein was effectively blocked in anti ZNRD1-SGC7901/VCRcells by gene transfection.ZNRD1 antisense nucleic acidtransfection sensitized drug resistant gastric cancer cells toVCR,increased ADM accumulation and inhibited the cellsproliferation.ZNRD1 antisense RNA transduction couldreverse the MDR of human drug-resistant gastric cancercell SGC7901/VCR to a degree.
AIM: To investigate the expression level of ZNRD1 gene ingastric cancer cells SGC7901 and gastric cancer MDR (multidrug resistant) cells SGC7901 / VCR, and to observe the drug sensitizing and proliferation effect of ZNRD1antisense nucleic acid transduction on SGC7901 / VCR cells. METHODS: Amplification of sequences encoding ZNRD1 from SGC7901 / VCR cDNA by PCR. The levels of ZNRD1 mRNA expression were demonstrated using semiquantitative reverse transcription polymerase chain reaction (RT-PCR). The Eukaryotic expression vector pcDNA3.1-anti ZNRD1 wasconstructed and transfected into SGC7901 / VCR cells bylipofectamine. Immunochemical method was used to detect the expression of protein in SGC7901 / VCR cells and transfectants. The cell cycle alteration and the intracellulariririmycin (ADM) accumulation were observed by FACS. Growth curve and drug sensitization of cells for vincristine (VCR) were analyzed with MTT assay .RESULTS: We cloned the open reading frame of full-lengthZNRD1. The expression of ZNRD1 showed higher in SGC7901 / VCR than in SGC7901 cells. The antisense ZNRD1 drug-resistant clones were selected after gene transfection. Immunochemical results showed that the expression level of ZNRD1 protein was lower in anti ZNRD1-SGC7901 / VCR cells than that in non-transfectants. Comparing to SGC7901 / VCR and pcDNA3.1-SGC7901 / VCR, anti ZNRD1-SGC7901 / V-Vode followed in G1 phase, with aconcomitant decrease of cell population in S phase. FACSalso suggested intracellular ADM accumulation increased 2fold in SGC7901 / VCR cells after transfected with antisenseZNRD1.MTT assay showed that transfectants cellsproliferation was lagged and more sensitive to VCR thannon-transfectants.CONCLUSION: ZNRD1 gene displayed highly expressionin VCR resistant gastric cancer cells. Expression of ZNRD1protein was effectively blocked in anti ZNRD1-SGC7901 / VCRcells by gene transfection.ZNRD1 antisense nucleic acidtransfection sensitized drug resistant gastric cancer cells toVCR, increased ADM accumulation and inhibited the cellsproliferation.ZNRD1 antisense RNA transduction couldreverse the MDR of human drug-resistant gastric cancer cell SGC7901 / VCR to a degree.