外泌体来源的 hsa_circ_0076987 分子在胃癌中的表达及其对胃癌细胞生物学行为的影响

Expression of exosome-derived hsa_circ_0076987 in gastric cancer and its effect on the biological behavior of gastric cancer cells

摘要

目的: 探究 hsa_circ_0076987 在胃癌细胞和胃癌血浆外泌体中的表达情况及其对胃癌细胞生物学功能的影响。方法: 通过 circRNA 芯片技术,检测 6 对胃癌患者与健康体检者血浆外泌体中 circRNA 的表达情况;采用实时荧光定量 PCR(qRT-PCR)检测 hsa_circ_0076987 在胃癌细胞和血浆外泌体中的表达情况,并结合临床病理资料,分析其临床相关性;采用受试者工作特征(ROC)曲线分析 hsa_circ_0076987 对胃癌的诊断价值;采用过表达质粒转染胃癌细胞,过表达 hsa_circ_0076987;通过 CCK-8 和克隆形成实验检测胃癌细胞增殖能力变化,Transwell 迁移和侵袭实验检测胃癌细胞迁移与侵袭能力变化。结果: qRT-PCR 结果显示,hsa_circ_0076987 在胃癌细胞和胃癌血浆外泌体中低表达(P 均<0.01),且与肿瘤神经浸润相关(P<0.05);ROC 曲线分析显示,hsa_circ_0076987 曲线下面积(AUC)为0.709,灵敏度为0.606,特异度为0.697,95%CI 为0.585~0.833(P<0.05);过表达 hsa_circ_0076987 可以抑制胃癌细胞的增殖、迁移与侵袭能力。结论: hsa_circ_0076987 在胃癌细胞及血浆外泌体中低表达且与胃癌发生发展有关,有望成为胃癌诊断的新型标志物与治疗靶点。

Abstract

Objective: To investigate the expression of hsa_circ_0076987 in gastric cancer cells and plasma exosomes, as well as its effects on the biological functions of gastric cancer cells. Methods: The expression of circRNAs in plasma exosomes of six pairs of gastric cancer patients and healthy individuals was detected by circRNA microarray technology. qRT-PCR was used to detect the expression of hsa_circ_0076987 in gastric cancer cells and plasma exosomes, and its relationship with clinicopathological characteristics was analyzed. The diagnostic value of hsa_circ_0076987 in gastric cancer was analyzed by receiver operating characteristic (ROC) curve. hsa_circ_0076987 was overexpressed in gastric cancer cells by transfection with overexpression plasmids. The changes in the cell proliferation, migration and invasion ability of gastric cancer cells were detected by CCK-8 assay, cell clone formation assay, Transwell migration and invasion assay, respectively. Results: The results of qRT-PCR showed that hsa_circ_0076987 was lowly expressed in gastric cancer cells and gastric cancer plasma exosomes (both P<0.01) and correlated with tumor neural infiltration (P<0.05). The ROC curve showed that the area under the curve (AUC) of hsa_circ_0076987 was 0.709, the sensitivity was 0.606, the specificity was 0.697, 95%CI was 0.585-0.833 (P<0.05). Overexpression of hsa_circ_0076987 inhibited the proliferation, migration and invasion ability of gastric cancer cells. Conclusion: hsa_circ_0076987 is lowly expressed in gastric cancer cells and plasma exosomes, and is associated with the development and progression of gastric cancer. It′s expected to be a novel marker and therapeutic target for gastric cancer diagnosis.

关键词

胃癌 / 外泌体 / hsa_circ_0076987 / 细胞增殖 / 细胞迁移 / 细胞侵袭

Key words

gastric cancer / exosome / hsa_circ_0076987 / cell proliferation / cell migration / cell invasion

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