核糖体蛋白S13(Ribosomal protein S13,RPS13)属于核糖体蛋白家族,在核糖体中的生物学功能主要是维持核糖体大小亚基之间的动态连接。从蛇床子[Cnidium monnieri(L.)Cuss.]的根、茎、叶组织中提取总RNA,通过RT-PCR扩增蛇床子核糖体蛋白s...核糖体蛋白S13(Ribosomal protein S13,RPS13)属于核糖体蛋白家族,在核糖体中的生物学功能主要是维持核糖体大小亚基之间的动态连接。从蛇床子[Cnidium monnieri(L.)Cuss.]的根、茎、叶组织中提取总RNA,通过RT-PCR扩增蛇床子核糖体蛋白s13(rps13)的开放阅读框(ORF),成功获得了一段453 bp的基因,该基因片段编码151个氨基酸的蛋白;将获得的目的基因rps13 ORF连接到表达载体p ET22b(+)中,并进一步转化到大肠杆菌表达菌中诱导目的基因体外表达;通过对蛋白结构进行分析表明蛇床子RPS13含有6个α螺旋;对蛇床子RPS13进行系统进化分析表明该蛋白在进化上具有高度保守性,与植物类胡萝卜的RPS13亲缘关系最近。蛇床子RPS13蛋白在体外的成功表达以及对其进行系统的生物信息学分析为进一步深入研究该蛋白的结构和功能奠定了理论基础。展开更多
Aims:Prevous study using differential display-PCR had found up-regulation of ribosomal protein S13(RPS13) and L23(RPL23) in vincristine-resistant gastric cancer cells.The aim of this study was to explore the associati...Aims:Prevous study using differential display-PCR had found up-regulation of ribosomal protein S13(RPS13) and L23(RPL23) in vincristine-resistant gastric cancer cells.The aim of this study was to explore the association of RPS13 and RPL23 with multidrug-resistant phenotype of gastric cancer cells.Methods:Northern blot analysis was used to determine the expression of RPS13 and RPL23 in vincristine-resistant gastric cancer cells SGC7901/VCR and its parental cells SGC7901.The full-length cDNAs encoding RPS13 and RPL23 were amplified from SGC7901/VCR cells using RT-PCR.Their sense and antisense expression vectors were constructed by DNA recombinant technique,and transferred into SGC7901 cells(sense vectors) or SGC7901/VCR cells(antisense vectors)by means of Lipofactamine.Drug sensitivity of gastric cancer cells was evalu-ated using MTT assay.Cell cycle analysis was performed using flow cytometry and proliferous index(PI) was calculated.Results:As Northem blot analysis indicated,RNA from SGC7902/VCR cells exhibited moderate signals of PRL23 and RPS13,while RNA from SGC7901 cells exhibited no signal of RPL23 and very weak signal of RPS13.RNA dot blot analysis indicated that RPS13 or RPL23 upregulated SGC7901 cells(SGC7901-RPS13,SGC7901-RPL23)and RPS13 or RPL23 down-regulated SGC7901/VCR cells(SGC7901/VCR-anRPS13,SGC7901/VCR-anRPL23) were successfully prepared by gene transduction.The results of MTT assay indicated that,comparing with non-transfected and empty vector transfected cells,SGC7901-RPL23 cells showed significantly increased IC50 values and resistance index(RI) of vincristine(VCR),adriamycin(ADR),5-fludrouracil(5-Fu) and mitomycin(MMC);SGC7901-RPS12 cells showed significantly increased IC50 values and RI of VCR,ADR and 5-Fu;SGC7901/VCR-anRPL23 cells showed significantly decreased IC50 values and RI of MMC and cisplatin(DDP);SGC7901/VCR-anRPS13 cells showed significantly decreased IC50 values and RI of VCR and MMC.Cell cycle analysis indicated that,comparing with non-transfected and empty vector transfected cells,SGC7901-RPS13 cells showed increased S phase and PI;SGC7901-anRPS13 cells and SGC7901-anRPL23 cells showed decreased S phase and PI.Conclusion:RPL23 and RPS13 are up-regulated in vincristine-resistant gastric cancer cells.Modulation of RPL23 and RPS13 could alter multidrug-resistant phenotype of gastric cancer cells.展开更多
文摘Aims:Prevous study using differential display-PCR had found up-regulation of ribosomal protein S13(RPS13) and L23(RPL23) in vincristine-resistant gastric cancer cells.The aim of this study was to explore the association of RPS13 and RPL23 with multidrug-resistant phenotype of gastric cancer cells.Methods:Northern blot analysis was used to determine the expression of RPS13 and RPL23 in vincristine-resistant gastric cancer cells SGC7901/VCR and its parental cells SGC7901.The full-length cDNAs encoding RPS13 and RPL23 were amplified from SGC7901/VCR cells using RT-PCR.Their sense and antisense expression vectors were constructed by DNA recombinant technique,and transferred into SGC7901 cells(sense vectors) or SGC7901/VCR cells(antisense vectors)by means of Lipofactamine.Drug sensitivity of gastric cancer cells was evalu-ated using MTT assay.Cell cycle analysis was performed using flow cytometry and proliferous index(PI) was calculated.Results:As Northem blot analysis indicated,RNA from SGC7902/VCR cells exhibited moderate signals of PRL23 and RPS13,while RNA from SGC7901 cells exhibited no signal of RPL23 and very weak signal of RPS13.RNA dot blot analysis indicated that RPS13 or RPL23 upregulated SGC7901 cells(SGC7901-RPS13,SGC7901-RPL23)and RPS13 or RPL23 down-regulated SGC7901/VCR cells(SGC7901/VCR-anRPS13,SGC7901/VCR-anRPL23) were successfully prepared by gene transduction.The results of MTT assay indicated that,comparing with non-transfected and empty vector transfected cells,SGC7901-RPL23 cells showed significantly increased IC50 values and resistance index(RI) of vincristine(VCR),adriamycin(ADR),5-fludrouracil(5-Fu) and mitomycin(MMC);SGC7901-RPS12 cells showed significantly increased IC50 values and RI of VCR,ADR and 5-Fu;SGC7901/VCR-anRPL23 cells showed significantly decreased IC50 values and RI of MMC and cisplatin(DDP);SGC7901/VCR-anRPS13 cells showed significantly decreased IC50 values and RI of VCR and MMC.Cell cycle analysis indicated that,comparing with non-transfected and empty vector transfected cells,SGC7901-RPS13 cells showed increased S phase and PI;SGC7901-anRPS13 cells and SGC7901-anRPL23 cells showed decreased S phase and PI.Conclusion:RPL23 and RPS13 are up-regulated in vincristine-resistant gastric cancer cells.Modulation of RPL23 and RPS13 could alter multidrug-resistant phenotype of gastric cancer cells.
