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目的了解HSP90β 反义核酸转染细胞中HSP90β蛋白的表达。方法通过lipofectamine介导将HSP90β反义核酸重组子pcDNA HSP90转染人胃癌细胞系SGC7901、人胃癌多药耐药细胞系SGC7901/VCR、人肝癌细胞系HCC7402及人食管癌细胞系Ec109。经用G418进行筛选 ,对筛选出的阳性克隆用Westernblot检测HSP90β蛋白的表达。结果pcDNA HSP90转染人胃癌细胞系SGC7901 ,人胃癌耐药细胞系SGC7901/VCR ,人肝癌细胞系HCC7402及人食管癌细胞系Ec109后 ,用G418筛选出的阳性克隆 ,分别被命名为:AH SGC7901 ,AH SGC7901/VCR ,AH HCC7402及AH Ec109。Westernblot检测结果表明 ,AH SGC7901,AH SGC7901/VCR ,AH HCC7402及AH Ec109表达的HSP90β蛋白低于其亲本细胞。结论HSP90β反义核酸可封闭HSP90βmRNA ,使pcDNA HSP90转染的细胞AH SGC7901 ,AH SGC7901/VCR ,AH HCC7402及AH Ec109表达的HSP90β蛋白减少 ,为研究HSP90下调对肿瘤细胞生物学活性的影响提供了实验材料。
Objective To understand the expression of HSP90β in HSP90β antisense RNA transfected cells. Methods Human gastric cancer cell line SGC7901, human gastric cancer multidrug resistance cell line SGC7901 / VCR, human hepatoma cell line HCC7402 and human esophageal carcinoma cell line Ec109 were transfected with lipofectamine mediated HSP90β antisense nucleic acid recombinant pcDNA HSP90. After screening with G418, the positive clones were screened by Western blot to detect the expression of HSP90β protein. Results After transfected with human gastric cancer cell line SGC7901, human gastric cancer drug resistant cell line SGC7901 / VCR, human hepatoma cell line HCC7402 and human esophageal cancer cell line Ec109 by pcDNA HSP90, the positive clones screened by G418 were named as AH SGC7901 , AH SGC7901 / VCR, AH HCC7402 and AH Ec109. The results of Western blot showed that HSP90β protein expressed by AH SGC7901, AH SGC7901 / VCR, AH HCC7402 and AH Ec109 was lower than that of their parental cells. Conclusion HSP90β antisense oligonucleotide can block HSP90βmRNA and decrease the expression of HSP90β in AH SGC7901, AH SGC7901 / VCR, AH HCC7402 and AH Ec109 transfected with pcDNA-HSP90, which provides an experimental study on the effect of HSP90 down-regulation on tumor cell biological activity material.