LINE-1ORF-1p对ERα阳性乳腺癌细胞的体外影响研究
In vitro effect of long interspersed nucleotide acids element-1 ORF-1p on estrogen receptorα of positive human breast cancer cells
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摘要: 目的 探讨人反转座子长散在重复序列编码蛋白1(LINE-1 ORF-1p)对ERα阳性乳腺癌细胞ZR75-1生长的影响及可能机理。 方法 利用Lipofectin 2000将LINE-1 ORF-1的表达载体及其空载体,siRNA及其空载体转染进入ERα阳性乳腺癌细胞ZR75-1细胞。使用MTT法测定LINE-1 ORF-1p对ERα阳性乳腺癌细胞ZR75-1生长的影响;利用Luciferase检测LINE-1 ORF-1p对ERα转录活性的影响。 结果 MTT法观察结果发现,LINE-1 Flag-ORF-1p能够显著促进ZR75-1细胞的生长(P=0.018)。LINE-1 ORF-1p siRNA表达载体能够抑制ZR75-1细胞的生长(P=0.022),其体外抑制率为E2-:25.51%,P=0.025,E2+:64.29%,P=0.021。LINE-1 Flag-ORF-1p能够升高ERα的转录活性(P=0.008)。LINE-1 ORF-1psiRNA表达载体能够降低ERα的转录活性(P=0.015), 结论 LINE-1 ORF-1p能够通过升高ERα的转录活性,促进乳腺肿瘤细胞ZR75-1生长。Abstract: Objective To study the effect of long interspersed nucleotide acids element-1(LINE-1) ORF-1p on proliferation of estrogen receptor α(ERα) positive human breast cancer ZR75-1 cells and its possible mechanism. Methods LINE-1 ORF-1 expression vector siRNA or empty vector,was transfected into ERα positive ZR75-1 cells with Lipofectin 2000.Effect of LINE-1 ORF-1p on proliferation of ERα positive ZR75-1 cells and transcriptional activity of ERα was detected by MTT and luciferase assay,respectively. Results MTT assay showed that LINE-1 Flag-ORF-1p promoted the proliferation of ERα positive ZR75-1 cells(P=0.018) while LINE-1 ORF-1p expression vector siRNA inhibited the proliferation of ZR75-1 cells(P=0.022) with an in vitro inhibition rate of 25.51% and 64.29% for E2-and E2+,respectively(P<0.05).LINE-1 Flag-ORF-1 increased the transcriptional activity of ERα(P=0.05) while LINE-1 ORF-1p expression vector siRNA reduced the transcriptional activity of ERα(P=0.015). Conclusion LINE-1 ORF-1p promotes the proliferation of ZR75-1 cells by increasing the transcription activity of ERα.
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