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RNA 干扰抑制 CD147 表达可降低癌症干细胞样 HT-29 细胞的增殖、侵袭能力并增加其化疗敏感性。

Inhibition of CD147 expression by RNA interference reduces proliferation, invasion and increases chemosensitivity in cancer stem cell-like HT-29 cells.

机构信息

Department of Life Sciences, Nanjing Normal University, Nanjing, Jiangsu, P.R. China.

Central Laboratory, Nanjing First Hospital, Nanjing Medical University, Nanjing, Jiangsu, P.R. China.

出版信息

Int J Oncol. 2015 Oct;47(4):1476-84. doi: 10.3892/ijo.2015.3138. Epub 2015 Aug 27.

DOI:10.3892/ijo.2015.3138
PMID:26316167
Abstract

The association between CD147 and cancer stem cells (CSCs) provides a new angle for cancer treatments. The aim of this study was to investigate the biological roles of CD147 in colorectal CSCs. The Oct4-green fluorescent protein (GFP) vector was used to isolate CSCs and pYr-mir30-shRNA was used to generate short hairpin RNA (shRNA) specifically for CD147. After RNA interference (RNAi), CD147 was evaluated by reverse transcription‑quantitative PCR and western blot analysis, and its biological functions were assessed by MTT and invasion assays. The results showed that the differentiation of isolated CSC-like HT-29 cells was blocked and these cells were highly positive for CD44 and CD147. RNAi-mediated CD147 silencing reduced the expression of CD147 at both mRNA and protein levels. Moreover, the activities of proliferation and invasion were decreased obviously in CSCs. Knockdown of CD147 increased the chemosensitivity of CSC-like cells to gemcitabine, cisplatin, docetaxel at 0.1, 1 and 10 µM respectively, however, there was no significant difference among the three groups to paclitaxel at 10 µM. In conclusion, these results suggest that CD147 plays an important role in colorectal CSCs and might be regarded as a novel CSC-specific targeted strategy against colorectal cancer.

摘要

CD147 与癌症干细胞(CSCs)之间的关联为癌症治疗提供了一个新的角度。本研究旨在探讨 CD147 在结直肠 CSCs 中的生物学作用。使用 Oct4-绿色荧光蛋白(GFP)载体分离 CSCs,并使用 pYr-mir30-shRNA 生成针对 CD147 的短发夹 RNA(shRNA)。在 RNA 干扰(RNAi)之后,通过逆转录-定量 PCR 和 Western blot 分析评估 CD147,并通过 MTT 和侵袭测定评估其生物学功能。结果表明,分离的 CSC 样 HT-29 细胞的分化被阻断,这些细胞高度表达 CD44 和 CD147。CD147 的 RNAi 介导的沉默降低了 CD147 在 mRNA 和蛋白质水平上的表达。此外,CSCs 的增殖和侵袭活性明显降低。CD147 的敲低增加了 CSC 样细胞对吉西他滨、顺铂、多西他赛在 0.1、1 和 10 μM 时的化学敏感性,然而,在 10 μM 时紫杉醇三组之间没有显著差异。总之,这些结果表明 CD147 在结直肠 CSCs 中发挥重要作用,可能被视为针对结直肠癌的新型 CSC 特异性靶向策略。

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