摘要
目的：通过转染ＨＳＰ９０β反义核酸重组子ｐｃＤＮＡ－ＨＳＰ９０于４株人消化系肿瘤细胞系，以建立ＨＳＰ９０β蛋白低调肿瘤细胞系，并研究ＨＳＰ９０β蛋白低调后细胞的生长情况。方法：用Ｌｉｐｏｆｅｃｔａｍｉｎｅ介导将ｐｅＤＮＡ－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及人食道癌细胞系Ｅｃ１０９，用Ｇ４１８进行筛选，用ＲＮＡ酶保护分析法鉴定ＨＳＰ９０β反义核酸的表达，用Ｗｅｓｔｅｒｎ ｂｌｏｔ法检测ＨＳＰ９０β蛋白的表达。用细胞生长曲线研究基因转染细胞的生长情况。结果：ｐｃＤＮＡ－ＨＳＰ９０转染细胞ＡＨ－ＳＧＣ７９０１，ＡＨ－ＳＧＣ７９０１／ＶＣＲ，ＡＨ－ＨＣＣ７４０２及ＡＨ－Ｅｃ１０９有ＨＳＰ９０β反义ＲＮＡ表达，这些细胞ＨＳＰ９０β蛋白表达低调。研究还发现这些细胞的生长受到不同程度的抑制，其中ＡＨ－ＳＧＣ７９０１／ＶＣＲ及ＡＨ－Ｅｃ１０９细胞受抑程度更明显。结论：ｐｃＤＮＡ－ＨＳＰ９０转染细胞ＡＨ－ＳＧＣ７９０１，ＡＨ－ＳＧＣ７９０１／ＶＣＲ，ＡＨ－ＨＣＣ７４０２及ＡＨ－Ｅｃ１０９ ＨＳＰ９０β蛋白表达降低，细胞的生长受到不同程度的抑制。
Objective: To set up HSP90β Protein down-regulated digestive tumor cell lines and study the influence of HSP90β to tumor cells.Methods:pcDNA-HSP90 of HSP90β anti-sense RNA reconstructs was transfected into SGC7901 of human gastric cancer cell line, SGC7901/VCR of MDR-type human gastric cancer cell line, HCC7402 of human hepatic cancer cell line and Ec109 of human esophageal cancer cell line by lipofectamine.Positive clones were selected by G418. Antisense RNA of HSP90β of the positive clones was detected by RNase protection assay and the expression of HSP90β protein was assayed by Western blot. Cell growth was studied by cell growth curve. Results:pcDNA-HSP90 transfected cells were named AH-SGC7901, AHSGC790l/VCR, AH-HCC7402 and AH-Ec109 respectively and there were antisense RNA of HSP90β in them. HSP90β protein in them was lower than in their parental cells.Cell growth was inhibited to different degrees,and the growth of AH-SGC790l/ VCR and AH-Ec109 cells were most greatly inhibited. Conclusion:The expression of HSP90β protein in AH-SGC7901, AHSGC790l/VCR, AH-HCC7402 and AH-Ec109 cell lines was decreased and the study also showed that decreasing the expression of HSP90β could inhibit tumor cell growth.
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
2000年第2期102-105,共4页
Chinese Journal of Cancer Biotherapy
基金
国家自然科学基金第!39570806号
国家杰出青年基金第!3952020号资助
关键词
HSP90β
基因转染
RNA酶保护
肿瘤细胞
HSP90β
anti-sense RNA
gene transfection
RNase protection assay
Western blot
