Department of General Surgery, Affiliated People's Hospital of Jiangsu University, Zhenjiang, Jiangsu, China (mainland).
Clinical Medicine College of Jiangsu University, Zhenjiang, Jiangsu, China (mainland).
Med Sci Monit. 2019 Jul 24;25:5482-5492. doi: 10.12659/MSM.915251.
BACKGROUND The recombinant avirulent Newcastle disease virus (NDV) LaSota strain expressing the rabies virus glycoprotein (rL-RVG) can induce much greater apoptosis than can NDV in gastric carcinoma cells, but the mechanisms involved remains unclear. MATERIAL AND METHODS The 2 gastric carcinoma cell lines were divided into the rL-RVG group, the NDV group, and the PBS group. MTT assay was used to detect and analyze cell viability. siRNA for alpha7-nAChR, alpha7-nAChR antagonist, or alpha7-nAChR agonist, AKT antagonist, and p-AKT agonist were used for pretreatment. The protein expressions of RVG, NDV, alpha7-nAChR, cleaved caspase-3, p-AKT, PI3K, Bcl-2, and Bax proteins were detected by Western blot assay. Immunofluorescence was used to detect expressions of alpha7-nAChR proteins. Light microscopy, flow cytometry, and TUNEL assay were used to assess apoptosis. RESULTS The results showed that 2 virus concentrations over 10³ dilution caused greater cell proliferation inhibition. rL-RVG treatment increased the expression of alpha7-nAChR, cleaved caspase-3, and Bax protein but decreased the expression of p-AKT, PI3K, and Bcl-2 protein. When the groups were pretreated with alpha7-nAChR antagonist, the alpha7-nAChR, cleaved caspase-3, and Bax protein expression increased, but the expression of p-AKT, PI3K, and Bcl-2 protein was clearly decreased. However, the results in the alpha7-nAChR agonist group were the opposite. When treated with the AKT antagonist, the result was the same as in the rL-RVG treatment group. The result in the AKT agonist group was the opposite of that in the AKT antagonist group. Compared with the NDV group, the results of light microscopy, FCM, and TUNEL assay showed that alpha7-nAChR antagonist significantly affected the apoptosis of gastric cancer cells in the rL-RVG group. CONCLUSIONS rL-RVG leads to much greater apoptosis through the alpha7-nAChR/PI3K/AKT pathway.
表达狂犬病病毒糖蛋白（rL-RVG）的重组弱毒新城疫病毒（NDV）LaSota 株在胃癌细胞中诱导的细胞凋亡比 NDV 多，但涉及的机制尚不清楚。
将 2 种胃癌细胞系分为 rL-RVG 组、NDV 组和 PBS 组。MTT 法检测和分析细胞活力。使用 α7-nAChR 的 siRNA、α7-nAChR 拮抗剂、α7-nAChR 激动剂、AKT 拮抗剂和 p-AKT 激动剂进行预处理。用 Western blot 法检测 RVG、NDV、α7-nAChR、cleaved caspase-3、p-AKT、PI3K、Bcl-2 和 Bax 蛋白的表达。用免疫荧光法检测 α7-nAChR 蛋白的表达。用光镜、流式细胞术和 TUNEL 法评估细胞凋亡。
结果表明，2 种病毒浓度超过 10³稀释度可引起更大的细胞增殖抑制。rL-RVG 处理增加了 α7-nAChR、cleaved caspase-3 和 Bax 蛋白的表达，但降低了 p-AKT、PI3K 和 Bcl-2 蛋白的表达。当用 α7-nAChR 拮抗剂预处理时，α7-nAChR、cleaved caspase-3 和 Bax 蛋白的表达增加，但 p-AKT、PI3K 和 Bcl-2 蛋白的表达明显减少。然而，在 α7-nAChR 激动剂组的结果则相反。用 AKT 拮抗剂处理时，结果与 rL-RVG 处理组相同。AKT 激动剂组的结果与 AKT 拮抗剂组相反。与 NDV 组相比，光镜、FCM 和 TUNEL 检测结果表明，α7-nAChR 拮抗剂显著影响 rL-RVG 组胃癌细胞的凋亡。
rL-RVG 通过 α7-nAChR/PI3K/AKT 通路导致更大的细胞凋亡。
