| 吴银霞
1
王业华
2
齐小康
2
顾沈阳
2
俞俊杰
2△.Let-7a 下调 k-Ras 和 c-Myc 癌基因的表达抑制肾癌细胞增殖[J].现代生物医学进展英文版,2014,14(6):1036-1039. |
| Let-7a 下调 k-Ras 和 c-Myc 癌基因的表达抑制肾癌细胞增殖 |
| Let-7a Inhibits the Proliferation in Renal Cell Carcinoma Cell Lines byRegulating k-Ras and c-Myc Gene Expression |
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| DOI： |
| 中文关键词: Let-7a KRAS c-myc 肾细胞癌 细胞增殖 |
| 英文关键词: Let-7a K-Ras C-Myc Renal cell cancer Cell proliferation |
| 基金项目: |
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| 中文摘要: |
| 摘要 目的： MicroRNA 是近年发现的一类单链小分子 RNA， 对它的研究已成为一个新的热点。最近的研究发现， 1et-7a 在细胞内
影响着基因的表达调控，在疾病发生中起着及极重要的作用， 尤其是在肿瘤的发展过程中， let-7a 扮演着不可替代的角色。本文主
要研究 let-7a 在肾癌细胞株中的表达情况及其调控的靶基因、 抑制细胞增殖的机制，对探索肾癌的致病基因， 寻求肾癌新的治疗
途径有重要意义。 方法： 应用化学合成的 let-7a 拟物（ mimics ）用脂质体 Lipofectamine 2000 在体外瞬时转染 786-O 和 Caki-1 肾
癌细胞株， 转染 48 小时后采用荧光定量 RT-PCR 的方法检测 let-7a 及 c-Myc、 k-Ras mRNA 的表达情况， Western blot 检测这两株
肾癌细胞转染了 let-7a mimics 后 c-Myc 及 k-Ras 蛋白的表达变化；转染 let-7a mimics 后分别在 24、 48、 72 小时三个时间点用
CCK-8 试剂盒检测对肾癌细胞株增殖的影响。 结果： 786-O 和 Caki-1 肾癌细胞株中 let-7a 的表达量明显低于正常肾小管上皮细胞
株 HK-2 （P<0.05 ） ; 转染了 let-7amimics 的 786-O 和 Caki-1 肾癌细胞株， RT-PCR 及 Western blot 结果显示 c-Myc、 k-Ras 在基因及
蛋白的表达水平明显下调 （P<0.05 ） ； CCK-8 检测结果显示转染了 let-7a mimics 的肾癌细胞株细胞增殖能力明显明显受到抑制， 与
阴性对照组比较差异有统计学意义 （P<0.05 ）。结论： Let-7a 在在肿瘤细胞与正常细胞中存在明显差异， let-7a 通过调控 c-Myc、
k-Ras 的表达能抑制肾癌细胞增殖。 Let-7a mimics 可以抑制肾癌细胞的增殖，因此上调 Let-7a 的表达有可能成为肾癌基因治疗的
一种有效治疗手段。 |
| 英文摘要: |
| ABSTRACT Objective:MicroRNA is a type of the single small RNA discovered in recent years, and the study of it has becomea
new hot spot. Let-7a plays a very important role and affects the expression of genes in cells. Especially let-7a plays an irreplaceable role
in the process of tumor development.This article mainly researches the expression of let-7a in kidney cancer cell lines and regulation of
target genes and the mechanism of inhibition of cell proliferation. It is a great significance to explore the pathogenic genes and new treatment approaches in kidney cancer.Methods: Application of chemical synthesis of the let-7a mimics with Lipofectamine 2000 transient
transfects in 786-O and Caki-1 kidney cancer cell lines was taken. The let-7a and c-Myc, k-Ras mRNA expressions were detected with
method of RT-PCR after transfection. The expression changeswere tested with Western blot . Effects of let-7a in cell proliferation were evaluated by CCK-8 assay.Results: Transfection of the let-7amimics in Cak-i and 786-Orenal cancer cell lines, that the c-Myc, k-Ras gene
and protein expression level significantly reduced(P<0.05). The expression levels of let-7a in 786-O and Caki-1 ells were lower than normal renal tubular epithelial cell lines HK-2 (P<0.05). Over-expression of let-7a in 786-O and Caki-1 cells suppressed c-Myc, k-Ras gene
and protein level. The result showed by RT-PCR and Western blot (P<0.05). CCK-8 test results showed that transfection let-7a mimics
inhibit their proliferation ability was significantly suppressed compared with negative control group(P<0.05).Conclusion； These findings
suggest that Let-7a plays an important role in kidney cancer cell lines proliferation through suppressing c-Myc, k-Ras in vitro , which providesa potential development of a new approach for the treatment of renal cancer. |
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