Silencing of miR-21 influences the function of colon cancer cell line HT-29 and the expression of PDCD4

任宇鹏; 宋纯; 张昊

doi:10.3969/j.issn.1007-3969.2015.01.002

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Silencing of miR-21 influences the function of colon cancer cell line HT-29 and the expression of PDCD4

更新时间：2025-12-31

- Silencing of miR-21 influences the function of colon cancer cell line HT-29 and the expression of PDCD4
- China Oncology Vol. 25, Issue 1, Pages: 6-12(2015)
- 作者机构：
  
  辽宁省肿瘤医院大肠外科,辽宁,沈阳,110042
- 作者简介：
- 基金信息：
- DOI：10.3969/j.issn.1007-3969.2015.01.002
  CLC：
- Published Online：08 May 2015，
  
  Published：08 May 2015
- 稿件说明：
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任宇鹏,宋纯,张昊. 下调miR-21对PDCD4表达及结肠癌HT-29细胞功能的影响研究[J]. 中国癌症杂志, 2015, 25(1): 6-12.

任宇鹏, 宋纯, 张昊. Silencing of miR-21 influences the function of colon cancer cell line HT-29 and the expression of PDCD4[J]. China Oncology, 2015, 25(1): 6-12.
任宇鹏,宋纯,张昊. 下调miR-21对PDCD4表达及结肠癌HT-29细胞功能的影响研究[J]. 中国癌症杂志, 2015, 25(1): 6-12. DOI： 10.3969/j.issn.1007-3969.2015.01.002.

任宇鹏, 宋纯, 张昊. Silencing of miR-21 influences the function of colon cancer cell line HT-29 and the expression of PDCD4[J]. China Oncology, 2015, 25(1): 6-12. DOI： 10.3969/j.issn.1007-3969.2015.01.002.

摘要

背景与目的：miR-21可能通过抑制PDCD4表达调控结肠癌浸润及转移等恶性行为。本研究通过下调miR-21表达后，检测结肠癌HT-29细胞功能的变化，并观察PDCD4在蛋白及mRNA表达水平的改变，探讨miR-21及PDCD4的表达在结肠癌恶性行为中的关系及机制。方法：构建靶向miR-21的干扰质粒simiR-21，转染HT-29细胞后，以实时定量PCR(qRT-PCR)法检测转染效率，MTT法检测转染后细胞增殖变化，流式细胞术检测转染后细胞调亡变化，Transwell检测迁移及浸润能力改变，蛋白质印迹法(Western blot)及qRT-PCR法检测干扰后PDCD4表达水平变化。结果：经qRT-PCR检测simiR-21在HT-29细胞转染效率为60%～65%，转染效率佳；MTT显示转染后72、96和120 h，HT-29增殖能力减弱(t=1.276，P0.05；t=3.276，P0.01；t=4.523，P0.01)；流式细胞术结果显示，与si-negative control及miR-21组对比转染后HT-29调亡率明显增加(t=2.132，P0.05；t=3.524，P0.05)；Transwell结果显示，simiR-21转染细胞迁移能力降低(t=2.423，P0.05；t=3.153，P0.05)，侵袭能力降低(t=3.245，P0.05；t=5.236，P0.05)；Western blot检测结果显示，PDCD4蛋白在simiR-21细胞的表达水平明显上调(t=2.342，P0.05；t=4.215，P0.05)；qRT-PCR检测结果显示，PDCD4 mRNA在simiR-21细胞的表达水平明显上调(t=2.261，P0.05；t=3.492，P0.05)。结论：simiR-21下调miR-21表达后，结肠癌HT-29细胞增殖能力受抑制，并促进其调亡，抑制迁移及浸润能力，PDCD4上调。miR-21可能通过下调PDCD4表达促进肿瘤细胞恶性行为，可作为结肠癌治疗新的靶向候选基因。

Abstract

Background and purpose: PDCD4 may be inhibited by miR-21 to regulate the malignant behaviors of colon cancer such as invasion and migration. This study aimed to explore the function of colon cancer HT-29 cell lines by downregulating miR-21 expression and discuss the mechanisms and relationship between miR-21 and PDCD4 in colon cancer malignant behaviors. Methods: simiR-21 was transfected into colon cancer cell line HT- 29 to downregulate the expression of miR-21. Proliferation

apoptosis

migration and invasion were detected by MTT

flow cytometry and Transwell assay after transfection. PDCD4 expression was detected by Western blot and qRTPCR. Results: The qRT-PCR analysis result proved that the transfection efficiency was 60%-65%. MTT analysis result showed that the proliferations of HT-29 cells were inhibited after the transfection of miR-21 for 72

120 h (t=1.276

P0.05; t=3.276

P0.01; t=4.523

P0.01). Comparing with si-negative control and miR-21 groups

flow cytometry result showed that the apoptosis rate was increased after miR-21 expression downregulated (t=2.132

P0.05; t=3.524

P0.05). Transwell assay result showed that migration (t=2.423

P0.05; t=3.153

P0.05) and invasion(t=3.245

P0.05; t=5.236

P0.05) were inhibited; Western blot result showed that PDCD4 expression was up-regulated at protein level(t=2.342

P0.05; t=4.215

P0.05); qRT-PCR result showed that PDCD4 expression was up-regulated at mRNA level(t=2.261

P0.05; t=3.492

P0.05). Conclusion: The proliferation

migration and invasion are the inhibited

and apoptosis is attenuated after miR-21 downregulated by simiR-21 transfection

PDCD4 expression is up- regulated. miR-21 may enhance the malignant behavior of cancer cells by downregulating the PDCD4 expression

miR- 21 might be a target gene for colon cancer therapy.

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