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HMGA2 表达的 RNA 干扰对 ACHN 肾癌细胞增殖和侵袭能力的影响。

Effect of RNA interference of the expression of HMGA2 on the proliferation and invasion ability of ACHN renal cell carcinoma cells.

机构信息

Xi'an Jiaotong University Health Science Center, Xi'an, Shanxi 710049, P.R. China.

Department of Urological Surgery, The Affiliated Zhongshan Hospital of Dalian University, Dalian, Liaoning 116001, P.R. China.

出版信息

Mol Med Rep. 2017 Oct;16(4):5107-5112. doi: 10.3892/mmr.2017.7258. Epub 2017 Aug 16.

Abstract

This aim of the present study was to observe the effect of high mobility group AT‑hook 2 (HMGA2) on the proliferation and invasion ability of ACHN renal cell carcinoma (RCC) cells. Human ACHN cells, an RCC cell line, and HKC normal human renal tubular epithelial cells were cultured. HMGA2 small interfering (si)RNA, Mock‑siRNA and their negative control group were designed and synthesized. Subsequently, the ACHN cells were transiently transfected using RNA interference technology. Finally, the mRNA and protein expression levels of HMGA2 were detected using reverse transcription-polymerase chain reaction and western blot analyses. The proliferation ability of the ACHN cells was determined using MTT, and ACHN cell invasion ability was detected using the Transwell method. The results showed that the mRNA and protein expression levels of HMGA2 in the ACHN cells were considerably higher, compared with those in the HKC cells (P<0.01). The RCC cells, in which the expression of HMGA2 was specifically silenced, was successfully constructed. The proliferation rate of cells in the HMGA2‑siRNA group was significantly lower, compared with that of cells in the Mock‑siRNA group and control group at 24, 48, 72 and 96 h post‑transfection (P<0.05). The invasion ability of cells in the HMGA2‑siRNA group was significantly lower, compared with that of cells in the Mock‑siRNA group and control group (P<0.05) 48 h following transfection. Therefore, the HMGA2 gene may function as an oncogene in the occurrence and development of RCC, and provide specific targets for the targeted therapy of RCC. Further detailed investigations of the HMGA2 gene are important for future gene therapy of RCC.

摘要

本研究旨在观察高迁移率族蛋白 A2(HMGA2)对 ACHN 肾细胞癌(RCC)细胞增殖和侵袭能力的影响。培养人 ACHN 细胞(一种 RCC 细胞系)和 HKC 正常人类肾小管上皮细胞。设计并合成了 HMGA2 小干扰(si)RNA、Mock-siRNA 及其阴性对照组。随后,使用 RNA 干扰技术瞬时转染 ACHN 细胞。最后,通过逆转录-聚合酶链反应和 Western blot 分析检测 HMGA2 的 mRNA 和蛋白表达水平。采用 MTT 法检测 ACHN 细胞的增殖能力,采用 Transwell 法检测 ACHN 细胞的侵袭能力。结果显示,与 HKC 细胞相比,ACHN 细胞中 HMGA2 的 mRNA 和蛋白表达水平明显升高(P<0.01)。成功构建了 HMGA2 表达特异性沉默的 RCC 细胞。HMGA2-siRNA 组细胞的增殖率在转染后 24、48、72 和 96 h 时明显低于 Mock-siRNA 组和对照组(P<0.05)。HMGA2-siRNA 组细胞的侵袭能力在转染后 48 h 时明显低于 Mock-siRNA 组和对照组(P<0.05)。因此,HMGA2 基因可能在 RCC 的发生发展中起癌基因作用,为 RCC 的靶向治疗提供了特定的靶点。进一步详细研究 HMGA2 基因对于未来 RCC 的基因治疗具有重要意义。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2630/5647043/42b871d1db53/MMR-16-04-5107-g00.jpg

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