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MicroRNA-184 通过下调 C-MYC 和 BCL-2 抑制人结肠癌细胞 SW480 和 HCT116 的增殖并促进其凋亡。

MicroRNA-184 inhibits proliferation and promotes apoptosis of human colon cancer SW480 and HCT116 cells by downregulating C-MYC and BCL-2.

机构信息

Department of General Surgery, Shanghai Pudong New Area People's Hospital, Shanghai, China.

Department of Neurology, Shanghai Pudong New Area People's Hospital, Shanghai, China.

出版信息

J Cell Biochem. 2018 Feb;119(2):1702-1715. doi: 10.1002/jcb.26330. Epub 2017 Sep 18.

Abstract

This study aimed to investigate the effects of microRNA-184 (miR-184) on the proliferation and apoptosis of human colon cancer cells through the regulation of C-MYC and BCL-2. Human colon cancer tissues were selected as case group, and adjacent normal tissues were as control group. Human colon cancer SW480 and HCT116 cells were allocated into blank, miR-184 mimic negative control (mimic-NC), miR-184 inhibitor NC (inhibitor-NC), miR-184 mimic, and miR-184 inhibitor groups. Flow cytometry, Annexin V/PI and MTT assay were used to examine the cell cycle, apoptosis and viability. The expressions of C-MYC, BCL-2 and miR-184 were detected via immunohistochemistry, Western blotting and reverse transcription quantitative polymerase chain reaction (RT-qPCR). C-MYC and BCL-2 were direct targets to miR-184. The growth of colon cancer cells in the miR-184 mimic group was inhibited and exhibited an increase in apoptosis. Cell growth in the miR-184 mimic group was increased in addition to the inhibition of apoptosis. Compared with miR-184 mimic group, the expressions of C-MYC and BCL-2 in miR-184 inhibitor group were increased. The expressions of C-MYC and BCL-2 in colon cancer tissues exhibited high levels of expression, while miR-184 displayed relatively low levels in comparison to the adjacent normal tissues. An association was detected regarding the expressions of miR-184, C-MYC and BCL-2 with the differentiation, invasion depth and lymph node metastasis. MiR-184 expression was negatively related to C-MYC and BCL-2 expressions. Our study suggested that miR-184 could inhibit proliferation and promote apoptosis of colon cancer cells by down-regulating expressions of C-MYC and BCL-2.

摘要

本研究旨在通过调节 C-MYC 和 BCL-2 来研究 microRNA-184(miR-184)对人结肠癌细胞增殖和凋亡的影响。选择人结肠癌组织作为病例组,选择相邻正常组织作为对照组。将人结肠癌 SW480 和 HCT116 细胞分为空白组、miR-184 模拟阴性对照组(mimic-NC)、miR-184 抑制剂 NC(inhibitor-NC)、miR-184 模拟组和 miR-184 抑制剂组。流式细胞术、Annexin V/PI 和 MTT 测定法用于检测细胞周期、细胞凋亡和细胞活力。通过免疫组织化学、Western blot 和逆转录定量聚合酶链反应(RT-qPCR)检测 C-MYC、BCL-2 和 miR-184 的表达。C-MYC 和 BCL-2 是 miR-184 的直接靶标。miR-184 模拟物组中结肠癌细胞的生长受到抑制,并且凋亡增加。miR-184 模拟物组中细胞生长增加,同时抑制了细胞凋亡。与 miR-184 模拟物组相比,miR-184 抑制剂组中 C-MYC 和 BCL-2 的表达增加。与相邻正常组织相比,结肠癌组织中 C-MYC 和 BCL-2 的表达水平较高,而 miR-184 的表达水平较低。miR-184、C-MYC 和 BCL-2 的表达与分化、浸润深度和淋巴结转移有关。miR-184 的表达与 C-MYC 和 BCL-2 的表达呈负相关。本研究表明,miR-184 可能通过下调 C-MYC 和 BCL-2 的表达来抑制结肠癌细胞的增殖并促进其凋亡。

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