Department of Oncology, Heilongjiang Province Hospital.
Department of Hematology, Harbin Medical University Cancer Hospital.
Anticancer Drugs. 2021 Oct 1;32(9):909-918. doi: 10.1097/CAD.0000000000001083.
The aim of this study is to explore the expression and mechanism of circ_0078607 on proliferation and apoptosis of gastric cancer. Real time PCR (RT-PCR) was performed to detect the expression of circ_0078607 in gastric cancer tumor tissues, plasma and cell lines. Cell viability was detected by cell counting Kit-8. Cell proliferation ability was assessed by cell cycle assay. The samples were analyzed by flow cytometry for the detection of apoptosis. Luciferase assay and RNA immunoprecipitation (RIP) were carried out to verify the relationship between circ_0078607 and miR-188-3p, miR-188-3p, and RAP1B. Western blot was employed to detect the protein level of RAP1B, ERK1/2 and AKT. In vivo, the effect of circ_0078607 on gastric cancer tumor growth was detected by lentivirus vector injection. Here, we found the increased level of circ_0078607 in gastric cancer tissues, gastric cancer patients plasma and cell lines. Knockdown of circ_0078607 could prevent proliferation and induce cell apoptosis in MKN-28 cells. Then we verified that circ_0078607 could interact with miR-188-3p by performed luciferase assay and RIP. Furthermore, we observed that RAP1B was a potential target of miR-188-3p. Next, we found that miR-188-3p inhibitor or overexpression of RAP1B could prevent the anti-tumor function of sh-circ_0078607. Silencing of circ_0078607 inhibited ERK1/2/AKT signal pathways via regulating miR-188-3p/RAP1B. In vivo, knockdown of circ_0078607 inhibited tumor growth. Knockdown of circ_0078607 inhibits the proliferation and induces apoptosis of gastric cancer via miR-188-3p/RAP1B signal pathway.
本研究旨在探讨 circ_0078607 对胃癌增殖和凋亡的表达及机制。采用实时荧光定量 PCR（RT-PCR）检测胃癌组织、血浆和细胞系中 circ_0078607 的表达。采用细胞计数试剂盒-8 检测细胞活力。通过细胞周期分析评估细胞增殖能力。通过流式细胞术分析检测细胞凋亡。通过荧光素酶报告和 RNA 免疫沉淀（RIP）实验验证 circ_0078607 与 miR-188-3p、miR-188-3p 和 RAP1B 的关系。采用 Western blot 检测 RAP1B、ERK1/2 和 AKT 的蛋白水平。体内实验采用慢病毒载体注射检测 circ_0078607 对胃癌肿瘤生长的影响。结果发现，胃癌组织、胃癌患者血浆和细胞系中 circ_0078607 水平升高。敲低 circ_0078607 可抑制 MKN-28 细胞增殖并诱导细胞凋亡。然后通过荧光素酶报告和 RIP 实验验证 circ_0078607 可与 miR-188-3p 相互作用。此外，我们观察到 RAP1B 是 miR-188-3p 的潜在靶标。接着发现 miR-188-3p 抑制剂或 RAP1B 的过表达可阻止 sh-circ_0078607 的抗肿瘤作用。沉默 circ_0078607 通过调节 miR-188-3p/RAP1B 抑制 ERK1/2/AKT 信号通路。体内实验中，敲低 circ_0078607 抑制肿瘤生长。circ_0078607 通过 miR-188-3p/RAP1B 信号通路抑制胃癌增殖并诱导细胞凋亡。
