lncRNA SOX21-AS1的表达对卵巢癌细胞增殖和转移能力的影响及机制研究

doi:10.11958/20220316

摘要/Abstract

摘要：

目的探讨长链非编码RNA SRY-box转录因子21反义RNA 1（lncRNA SOX21-AS1）对卵巢癌细胞增殖和转移能力的影响及其作用机制。方法选取卵巢癌组织和其邻近的癌旁组织各75例，另选取卵巢癌细胞株SKOV3、A2780、OVCR3和人卵巢正常上皮细胞HOSE作为实验对象，实时荧光定量PCR法检测lncRNA SOX21-AS1在组织和细胞中的表达。根据是否干扰lncRNA SOX21-AS1表达将细胞分为si-NC组和si-SOX21-AS1组。MTS实验和平板克隆实验检测细胞增殖能力，Transwell实验检测细胞转移能力，TOP/FOP荧光素酶实验和Western blot实验检测细胞中Wnt/β-catenin信号通路活性和相关蛋白表达水平。结果与癌旁组织相比，lncRNA SOX21-AS1在卵巢癌组织中的表达上调（P<0.05）。与卵巢正常上皮细胞HOSE相比，lncRNA SOX21-AS1在卵巢癌细胞中的表达上调，SKOV3中最高（P<0.05）。与FIGO分期Ⅰ-Ⅱ期、无淋巴结转移的患者相比，FIGO分期Ⅲ-Ⅳ期、有淋巴结转移患者lncRNA SOX21-AS1相对表达水平升高（P<0.05）。干扰lncRNA SOX21-AS1表达可抑制卵巢癌细胞增殖和转移能力、抑制Wnt/β-catenin信号通路活性并降低Wnt/β-catenin信号通路各蛋白的表达水平（P<0.05）。结论 lncRNA SOX21-AS1在卵巢癌组织和细胞中高表达，干扰lncRNA SOX21-AS1的表达可抑制卵巢癌细胞增殖和转移能力。

关键词: 卵巢肿瘤, 细胞增殖, 肿瘤转移, 基因表达调控，肿瘤, 长链非编码RNA SRY-box转录因子21反义RNA 1

Abstract:

Objective To study the effect of long non-coding RNA SRY-box transcription factor 21 antisense RNA 1 (lncRNA SOX21-AS1) on the proliferation and metastasis of ovarian cancer cells and its mechanism. Methods A total of 75 samples of ovarian cancer and 75 samples of adjacent paracancerous tissues were selected. Ovarian cancer cell lines SKOV3, A2780, OVCR3 and human normal ovarian epithelial cells HOSE were selected as the experimental objects. The expression levels of lncRNA SOX21-AS1 in tissues and cells were detected by qPCR. The cells were divided into the si-NC group and the si-SOX21-AS1 group according to whether they interfered with the expression of lncRNA SOX21-AS1. MTS assay and plate cloning assay were used to detect cell proliferation ability, and Transwell assay was used to detect cell transfer ability. TOP/FOP luciferase assay and Western blot assay were used to detect Wnt/β-catenin signaling pathway activity and related protein expression levels in cells. Results Compared with adjacent tissues, the expression of lncRNA SOX21-AS1 was significantly up-regulated in ovarian cancer tissues (P<0.05). Compared with normal ovarian epithelial cells HOSE, the expression of lncRNA SOX21-AS1 was significantly up-regulated in ovarian cancer cells (P<0.05). Compared with patients with FIGO stage I-II and patients with no lymph node metastasis, the relative expression level of lncRNA SOX21-AS1 was significantly higher in patients with FIGO stage III-IV and patients with lymph node metastasis (P<0.05). Interfering with the expression of lncRNA SOX21-AS1 significantly inhibited cell proliferation and metastasis, Wnt/β-catenin signaling pathway activity and Wnt/β-catenin signaling pathway related protein expression (P<0.05). Conclusion lncRNA SOX21-AS1 is highly expressed in ovarian cancer tissues and cells. Interfering with the expression of lncRNA SOX21-AS1 significantly inhibits the proliferation and metastasis of ovarian cancer cells.

Key words: ovarian neoplasms, cell proliferation, neoplasm metastasis, gene expression regulation, neoplastic, lncRNA SOX21-AS1

中图分类号:

R737.31

图/表 5

表1 不同临床特征的卵巢癌患者lncRNA SOX21-AS1相对表达水平比较

Tab.1 Comparison of relative expression levels of lncRNA SOX21-AS1 between ovarian cancer patients with different clinical features

临床特征	n	lncRNA SOX21-AS1相对表达量	t
年龄（岁）
≤60	45	1.65±0.31	0.658
>60	30	1.70±0.34	0.658
组织分级
G1-2	34	1.61±0.29	1.436
G3	41	1.72±0.36	1.436
FIGO分期
Ⅰ-Ⅱ	35	1.45±0.24	5.193^＊＊
Ⅲ-Ⅳ	40	1.83±0.37	5.193^＊＊
淋巴结转移
有	44	1.79±0.34	3.905^＊＊
无	31	1.50±0.28	3.905^＊＊
远处转移
有	28	1.75±0.34	1.829
无	47	1.62±0.27	1.829

表2 2组细胞lncRNA SOX21-AS1表达、增殖、转移和Wnt/β-catenin信号通路活性的比较（n=3，$\bar{x}\pm s$）

Tab.2 Comparison of lncRNA SOX21-AS1 expression, proliferation, transfer and Wnt/β-catenin signaling pathway activity between two groups of cells

组别	lncRNA SOX21-AS1相对表达量	SKOV3增殖能力（A₄₉₀）			克隆团形成数（个/视野）	细胞转移数目（个/视野）	TOP/FOP荧光素酶活性
组别	lncRNA SOX21-AS1相对表达量	24 h	48 h	72 h	克隆团形成数（个/视野）	细胞转移数目（个/视野）	TOP/FOP荧光素酶活性
si-NC组	0.98±0.02	0.55±0.05	0.81±0.05	1.30±0.07	70.67±6.81	92.33±12.54	0.97±0.03
si-SOX21-AS1组	0.38±0.07	0.47±0.04	0.52±0.03	0.73±0.04	51.33±4.32	50.67±8.56	0.54±0.08
t	14.275^＊＊	2.164	8.614^＊＊	12.246^＊＊	4.154^＊	4.752^＊	8.717^＊＊

图1 干扰lncRNA SOX21-AS1的表达对卵巢癌细胞的增殖、转移能力的影响 A：2组细胞的克隆团形成数比较（结晶紫染色，×1）；B：2组转移能力比较（结晶紫染色，×100）。

Fig.1 The effect of interfering the expression of lncRNA SOX21-AS1 on the proliferation and metastasis of ovarian cancer cells

表3 2组细胞Wnt/β-catenin信号通路相关蛋白表达比较（n=3，$\bar{x}\pm s$）

Tab.3 Comparison of Wnt/β-catenin signaling pathway related protein expression between the two groups of cells

组别	Wnt3a	β-catenin	CyclinD1	cMyc	MMP-7
si-NC组	0.91±0.06	0.47±0.03	0.52±0.06	1.18±0.14	0.98±0.10
si-SOX21-AS1组	0.36±0.05	0.31±0.02	0.11±0.03	0.19±0.02	0.27±0.02
t	12.197^＊＊	7.686^＊	10.586^＊＊	12.125^＊＊	12.059^＊＊

图2 干扰lncRNA SOX21-AS1对卵巢癌细胞Wnt/β-catenin信号通路相关蛋白的表达影响

Fig.2 Effects of interfering lncRNA SOX21-AS1 on the expression of Wnt/β-catenin signaling pathway related proteins in ovarian cancer cells

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