[摘要] 目的: 探讨长链非编码RNA（lncRNA）FOXD2-AS1 是否通过靶向miR-506-5p 调控宫颈癌细胞增殖和凋亡。方法:体外培养人宫颈癌细胞系HeLa、Siha、Caski 与正常宫颈细胞株Ect1/E6E7，用qPCR检测细胞中FOXD2-AS1 和miR-506-5p 的表达水平。用脂质体转染技术分别构建抑制FOXD2-AS1 表达和过表达miR-506-5p 的宫颈癌细胞，用MTT实验和流式细胞术检测细胞的增殖和凋亡情况，WB实验检测细胞中增殖相关蛋白CyclinD1、p21 和p27 及凋亡相关蛋白Bcl-2、BAX和cleaved-capase-3的表达。用双荧光素酶报告基因实验验证FOXD2-AS1 是否靶向miR-506-5p，并分析同时抑制FOXD2-AS1 和miR-506-5p 表达对宫颈癌细胞增殖与凋亡的影响。结果：与Ect1/E6E7 细胞比较，宫颈癌HeLa、Siha 和Caski 细胞中FOXD2-AS1 表达水平显著升高，miR-506-5p 表达水平降低（均P<0.01）。抑制FOXD2-AS1 表达可显著抑制宫颈癌细胞中CyclinD1 蛋白和Bcl-2 蛋白表达，并促进p21、p27 和BAX、cleaved-capase-3 蛋白的表达，抑制细胞增殖并促进细胞凋亡（均P<0.01）。过表达miR-506-5p 可显著抑制宫颈癌细胞中CyclinD1 和Bcl-2 蛋白表达，促进p21 和BAX蛋白表达，抑制细胞增殖并促进细胞凋亡（均P<0.01）。双荧光素酶报告基因实验证实宫颈癌细胞中FOXD2-AS1 靶向负调控miR-506-5p 的表达（P<0.01）。抑制miR-506-5p 表达逆转了抑制FOXD2-AS1 表达对宫颈癌细胞增殖和凋亡的作用（P<0.01）。结论: FOXD2-AS1 通过靶向miR-506-5p 的表达调控宫颈癌细胞的增殖和凋亡。

[Abstract] Objective: To investigate whether long non-coding RNA (lncRNA) FOXD2-AS1 targets miR-506-5p to regulate proliferation and apoptosis of cervical cancer cells. Methods: Human normal cervical cells Ect1/E6E7 and cervical cancer cell lines (HeLa, Siha and Caski) were cultured in vitro, and the expression levels of FOXD2-AS1 and miR-506-5p in cells were detected by qPCR. The cervical cancer cells with FOXD2-AS1 knockdown and miR-506-5p over-expression were constructed by liposome transfection technology, and the proliferation and apoptosis of cells were detected by MTT assay and flow cytometry respectively, the expression of proliferation-related proteins CyclinD1, p21, p27 and apoptosis-related proteins Bcl-2, BAX, cleaved-capase-3 were detected by WB. Dual luciferase reporter assay was used to verify whether FOXD2-AS1 would target miR-506-5p; and the effects of simultaneous inhibition of FOXD2-AS1 and miR-506-5p on proliferation and apoptosis of cervical cancer cells were also analyzed. Results: Compared with Ect1/E6E7 cells, the expression of FOXD2-AS1 significantly increased while the expression of miR-506-5p significantly decreased in cervical cancer HeLa, Siha and Caski cells (all P<0.01). FOXD2-AS1 knockdown significantly inhibited the protein expressions of CyclinD1, Bcl-2 and cell proliferationin cervical cancer cells, but promoted the protein expressions of p21, p27, BAX, cleavedcapase-3, and cell apoptosis (all P<0.01). miR-506-5p over-expression significantly inhibited the protein expressions of CyclinD1, Bcl-2 and cell proliferation in cervical cancer cells, but promoted the protein expressions of p21, BAX, and cell apoptosis (all P<0.01).Dual luciferase reporter gene assay confirmed that FOXD2-AS1 negatively regulated the expression of miR-506-5p in cervical cancer cells (P<0.01). Inhibition of miR-506-5p expression reversed the effect of FOXD2-AS1 knockdown on proliferation and apoptosis of cervical cancer cell (P<0.01). Conclusion: FOXD2-AS1 modulates proliferation and apoptosis of cervical cancer cells by negatively regulating the expression of miR-506-5p.

广东省中医药局科研项目（No.20173031）