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miR-375 通过靶向 sp1 抑制结直肠癌细胞增殖。

The inhibitory effect of miR-375 targeting sp1 in colorectal cancer cell proliferation.

机构信息

Department of Oncology, Xiangyang No. 1 People's Hospital, Hubei University of Medicine, Xiangyang, Hubei, China.

出版信息

Eur Rev Med Pharmacol Sci. 2018 Jan;22(2):405-411. doi: 10.26355/eurrev_201801_14188.

Abstract

OBJECTIVE

Sp1 is a member of super zinc finger structure family that participates in cancer cells' apoptosis, proliferation, survival, and differentiation. This study detected the expressions of miR-375 and sp1 in colorectal cancer tissue and cells to analyze their impact on cell proliferation.

PATIENTS AND METHODS

Colorectal cancer patients in our hospital were enrolled. HCT-116 cell was transfected with miR-375 mimics, mimics control, and miR-375 + sp1, respectively. RT-PCR and Western blot were applied to detect expressions of miR-375 and sp1 at mRNA and protein level in colorectal cancer tissue, para-carcinoma tissue, and normal colorectal tissue. RT-PCR and Western blot were used to test levels of miR-375 and sp1 in HCT-116 cells after transfection. MTT assay was performed to determine HCT-116 cell proliferation.

RESULTS

Our data showed that miR-375 was downregulated, while sp1 was overexpressed in colorectal cancer tissue compared with that in para-carcinoma tissue and normal control (p < 0.05). MiR-375 level was elevated, while sp1 mRNA was declined after miR-375 mimic transfection (p < 0.05). Compared with miR-375 mimic group, the levels of miR-375 and sp1 showed no difference in miR-375 + sp1 group (p > 0.05). Of note, the increase of MiR-375 and reduction of sp1 were in a time-dependent manner (p < 0.05). The cell proliferation rate in miR-375 mimic group was significantly decreased compared with that in mimic control and blank group (p < 0.05). The cell proliferation rate in miR-375 + sp1 group was significantly higher than that miR-375 group, but still lower than the control (p < 0.05). The proliferation rate gradually declined in a time-dependent manner (p < 0.05).

CONCLUSIONS

MiR-375 was decreased and sp1 level was enhanced in colorectal cancer. MiR-375 suppresses the proliferation of colorectal cancer cells via the inhibition of sp1 expression at posttranscriptional level.

摘要

目的

Sp1 是超级锌指结构家族的成员,参与癌细胞的凋亡、增殖、存活和分化。本研究检测了结直肠癌组织和细胞中 miR-375 和 sp1 的表达,分析它们对细胞增殖的影响。

患者和方法

选取我院收治的结直肠癌患者。分别将 HCT-116 细胞转染 miR-375 模拟物、模拟物对照和 miR-375+sp1。应用 RT-PCR 和 Western blot 检测结直肠癌组织、癌旁组织和正常结直肠组织中 miR-375 和 sp1 的表达。应用 RT-PCR 和 Western blot 检测转染后 HCT-116 细胞中 miR-375 和 sp1 的水平。MTT 法检测 HCT-116 细胞的增殖情况。

结果

数据显示,与癌旁组织和正常对照组相比,结直肠癌组织中 miR-375 表达下调,sp1 表达上调(p<0.05)。转染 miR-375 模拟物后 miR-375 水平升高,sp1 mRNA 水平下降(p<0.05)。与 miR-375 模拟物组相比,miR-375+sp1 组 miR-375 和 sp1 水平无差异(p>0.05)。值得注意的是,miR-375 的增加和 sp1 的减少呈时间依赖性(p<0.05)。miR-375 模拟物组细胞增殖率明显低于模拟物对照组和空白组(p<0.05)。miR-375+sp1 组细胞增殖率明显高于 miR-375 组,但仍低于对照组(p<0.05)。增殖率呈时间依赖性逐渐下降(p<0.05)。

结论

结直肠癌中 miR-375 降低,sp1 水平升高。miR-375 通过抑制 sp1 的转录后表达抑制结直肠癌细胞的增殖。

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