Department of Cardiothoracic Surgery, Tianjin Medical University General Hospital, Tianjin, China.
Eur Rev Med Pharmacol Sci. 2020 May;24(10):5404-5411. doi: 10.26355/eurrev_202005_21324.
The aim of this study was to investigate the expression of long non-coding ribonucleic acid (lncRNA) AK058003 in esophageal carcinoma (EC) tissues, and to analyze its intervention effect.
The expression of lncRNA AK058003 in EC tissues and para-carcinoma tissues from 130 EC patients was detected via quantitative Polymerase Chain Reaction (qPCR). EC cell lines were selected for exogenous interference in lncRNA AK058003. Subsequently, the expression of lncRNA AK058003 in normal esophageal epithelial cell line (Het-1A) and EC cell lines (EC109, EC9706, KYSE-150, KYSE-30, and TE-1) was detected by qPCR. EC9706 cell lines with the highest expression of lncRNA AK058003 were selected and transfected with lncRNA AK058003 siRNA and lncRNA AK058003 control, respectively. After transfection, the expression of lncRNA AK058003 was determined using PCR. The changes in cell growth and proliferation were analyzed via cell growth curve and cell cycle assay. Meanwhile, the changes in cell migration and invasion were analyzed through wound healing assay. Protein expressions of matrix metalloproteinase-1 (MMP1) and MMP2 were determined by Western blot. Clinical data were collected from EC patients, and the association between lncRNA AK058003 expression and tumor-node-metastasis (TNM) stage was finally analyzed.
LncRNA AK058003 was highly expressed EC tissues compared with para-carcinoma tissues (p<0.01). Compared with Het-1A cells, the expression of lncRNA AK058003 was significantly higher in EC109, EC9706, KYSE-150, KYSE-30, and TE-1 cells, with highest level in EC9706 cells (p<0.05). The expression of lncRNA AK058003 remarkably declined in lncRNA AK058003 siRNA group compared with lncRNA AK058003 control group (p<0.001). Compared with lncRNA AK058003 control group, the proliferation of EC cells was significantly weakened in lncRNA AK058003 siRNA group, with the greatest difference at 3 d. Flow cytometry results revealed that cell cycle was arrested in G0/G1 phase in lncRNA AK058003 siRNA group. Wound healing assay indicated that the intercellular distance became large, and cell migration ability was evidently enhanced in lncRNA AK058003 siRNA group with time (p<0.05). Besides, the protein expressions of MMP1 and MMP2 were remarkably lower in lncRNA AK058003 siRNA group than those in lncRNA AK058003 control group. This indicated remarkably declined invasion and metastasis ability. In addition, the postoperative prognosis was significantly worse in patients with higher expression of lncRNA AK058003 (p<0.05). All these findings suggested that lncRNA AK058003 could serve as a biomarker for EC prognosis.
LncRNA AK058003 is highly expressed in EC patients, which promotes proliferation, migration, invasion, and metastasis of EC cells. In addition, the postoperative prognosis of EC patients with high expression of lncRNA AK058003 is relatively poor.
本研究旨在探讨长链非编码 RNA(lncRNA)AK058003 在食管癌(EC)组织中的表达,并分析其干预作用。
采用实时定量聚合酶链反应(qPCR)检测 130 例 EC 患者癌组织及癌旁组织中 lncRNA AK058003 的表达。选择 EC 细胞系进行 lncRNA AK058003 的外源性干扰。随后,采用 qPCR 检测正常食管上皮细胞系(Het-1A)和 EC 细胞系(EC109、EC9706、KYSE-150、KYSE-30 和 TE-1)中 lncRNA AK058003 的表达。选择 lncRNA AK058003 表达最高的 EC9706 细胞系,分别转染 lncRNA AK058003 siRNA 和 lncRNA AK058003 对照,转染后采用 PCR 检测 lncRNA AK058003 的表达。通过细胞生长曲线和细胞周期分析观察细胞生长和增殖的变化。同时,通过划痕愈合试验分析细胞迁移和侵袭的变化。采用 Western blot 检测基质金属蛋白酶-1(MMP1)和 MMP2 的蛋白表达。收集 EC 患者的临床资料,分析 lncRNA AK058003 表达与 TNM 分期的相关性。
lncRNA AK058003 在 EC 组织中的表达明显高于癌旁组织(p<0.01)。与 Het-1A 细胞相比,lncRNA AK058003 在 EC109、EC9706、KYSE-150、KYSE-30 和 TE-1 细胞中的表达明显升高,在 EC9706 细胞中表达最高(p<0.05)。与 lncRNA AK058003 对照组相比,lncRNA AK058003 siRNA 组 lncRNA AK058003 的表达明显下降(p<0.001)。与 lncRNA AK058003 对照组相比,lncRNA AK058003 siRNA 组 EC 细胞的增殖能力明显减弱,第 3 天差异最大。流式细胞术结果显示,lncRNA AK058003 siRNA 组细胞周期停滞在 G0/G1 期。划痕愈合试验表明,lncRNA AK058003 siRNA 组随着时间的推移细胞间距离增大,细胞迁移能力明显增强(p<0.05)。此外,lncRNA AK058003 siRNA 组 MMP1 和 MMP2 的蛋白表达明显低于 lncRNA AK058003 对照组。这表明侵袭和转移能力明显下降。此外,lncRNA AK058003 高表达的 EC 患者术后预后明显较差(p<0.05)。所有这些发现表明,lncRNA AK058003 可作为 EC 预后的生物标志物。
lncRNA AK058003 在 EC 患者中高表达,促进 EC 细胞的增殖、迁移、侵袭和转移。此外,lncRNA AK058003 高表达的 EC 患者术后预后较差。