重组新城疫病毒rL-RVG抑制人肺腺癌细胞系增殖

doi:10.16352/j.issn.1001-6325.2023.10.1549

基础医学与临床 ›› 2023, Vol. 43 ›› Issue (10): 1549-1556.doi: 10.16352/j.issn.1001-6325.2023.10.1549

重组新城疫病毒rL-RVG抑制人肺腺癌细胞系增殖

张珍珍¹, 李洋², 高生宝², 夏德鑫², 严玉兰^1,3*

1.江苏大学附属人民医院呼吸内科,江苏镇江 212000;
2.江苏大学医学院临床医学系,江苏镇江 212000;
3.香港大学深圳医院呼吸内科,广东深圳 518000

收稿日期:2023-02-02 修回日期:2023-06-12 出版日期:2023-10-05 发布日期:2023-10-05
通讯作者: *yulanyan@163.com
基金资助:
镇江市社会发展项目(SH2021032)

Recombinant Newcastle virus rL-RVG inhibits proliferation of human lung adenocarcinoma cell lines

ZHANG Zhenzhen¹, LI Yang², GAO Shengbao², XIA Dexin², YAN Yulan^{1, 3*}

1. Department of Respiratory Medicine, People's Hospital Affiliated to Jiangsu University, Zhenjiang 212000;
2. Department of Clinical Medicine, School of Medicine, Jiangsu University, Zhenjiang 212000;
3. Department of Respiratory Medicine, the University of Hong Kong-Shenzhen Hospital, Shenzhen 518000,China

Received:2023-02-02 Revised:2023-06-12 Online:2023-10-05 Published:2023-10-05
Contact: *yulanyan@163.com

摘要/Abstract

摘要： 目的探讨表达狂犬病毒糖蛋白的重组新城疫病毒rL-RVG抑制人肺腺癌细胞系A549和PC9增殖。方法体外培养人肺腺癌细胞系A549、PC9,将处于对数增殖期的细胞分为对照组、新城疫病毒感染组(NDV)、重组新城疫病毒感染组(rL-RVG)、铁死亡诱导剂组(Erastin)和重组新城疫病毒联合铁死亡诱导剂组(rL-RVG+Erastin)。病毒及药物感染细胞24 h后,光学显微镜下观察细胞形态,CCK-8法检测细胞增殖能力,划痕试验检测细胞迁移能力,细胞毒性检测试剂盒测定乳酸脱氢酶(LDH)释放量,流式细胞术检测细胞内活性氧(ROS)含量,Western blot检测铁死亡相关蛋白p53、SLC7A11、GPX4的表达。结果与对照组相比,NDV组、rL-RVG组、Erastin组细胞数、细胞增殖能力及迁移的距离明显降低(P均＜0.001),LDH释放量和总ROS水平显著提高(P＜0.01),p53蛋白表达明显增加(P＜0.001),而SLC7A11、GPX4蛋白表达明显降低(P＜0.001)。rL-RVG组与NDV组相比上述结果更明显(P＜0.05),rL-RVG+Erastin组与rL-RVG组和Erastin组相比细胞数、细胞增殖能力及迁移的距离明显减少(P＜0.05),LDH释放量和总ROS水平显著提高(P＜0.05),P53蛋白表达明显增加(P＜0.001),而SLC7A11、GPX4蛋白表达明显降低(P＜0.001)。结论 rL-RVG可以发挥类似Erastin抑制肿瘤细胞增殖的作用,且其效果远远强于NDV。这为rL-RVG诱导的细胞死亡提供了新的见解,并强调了病毒在治疗肿瘤中的关键作用。

关键词: 重组新城疫病毒, 新城疫病毒, Erastin, 细胞增殖, 肺腺癌

Abstract: Objective Exploration of recombinant Newcastle disease virus with stable expression of rabies virus glycoprotein(rL-RVG) to inhibit the proliferation of human lung adenocarcinoma cell lines A549 and PC9. Methods Human lung adenocarcinoma cell lines A549 and PC9 were cultured in vitro, and cells in logarithmic proliferation phase were collected and divided into control group, Newcastle disease virus infection group (NDV), recombinant Newcastle disease virus infection group(rL-RVG),ferroptosis group (Erastin) and recombinant Newcastle disease virus combined with ferroptosis inducer group (rL-RVG+Erastin). After 24 h of incubation, cell morphology was observed by microscope and cell proliferation was detected by CCK-8 method, cell migration was detected by scratch test, lactate dehydrogenase (LDH) release was measured by cytotoxicity assay kit, intracellular reactive oxygen species (ROS) content was detected by flow cytometry, and the expression of ferroptosis related proteins (p53, SLC7A11, GPX4) was detected by Western blot. Results Compared with the control group, the cell counting, cell proliferation and distance of migration were significantly reduced in the NDV group, rL-RVG group and Erastin group(P＜0.001). LDH release and total ROS level were significantly increased (P＜0.01), p53 protein expression was significantly increased(P＜0.001), while SLC7A11 and GPX4 protein expression were significantly decreased (P＜0.001). rL-RVG group had a more pronounced effect as compared with NDV group. rL-RVG+Erastin group had significantly decreased cell number, cell proliferation capacity and distance of migration compared with rl-RVG and Erastin groups(P＜0.05), and LDH release and total ROS level were significantly increased(P＜0.05), p53 protein expression was significantly increased (P＜0.001), while SLC7A11 and GPX4 protein expression were significantly decreased (P＜0.001). Conclusions rL-RVG can exert a similar effect to Erastin to inhibit tumor cell proliferation, and its effect is much stronger than that of NDV. This provides new insights into rL-RVG-induced cell death and highlights the critical role of oncolytic viruses in the treatment of tumors.

Key words: Newcastle disease virus(rL-RVG), Newcastle disease virus(NDV), Erastin, cell proliferation, lung adenocarcinoma

中图分类号:

R734.2

张珍珍, 李洋, 高生宝, 夏德鑫, 严玉兰. 重组新城疫病毒rL-RVG抑制人肺腺癌细胞系增殖[J]. 基础医学与临床, 2023, 43(10): 1549-1556.

ZHANG Zhenzhen, LI Yang, GAO Shengbao, XIA Dexin, YAN Yulan. Recombinant Newcastle virus rL-RVG inhibits proliferation of human lung adenocarcinoma cell lines[J]. Basic & Clinical Medicine, 2023, 43(10): 1549-1556.

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重组新城疫病毒rL-RVG抑制人肺腺癌细胞系增殖

Recombinant Newcastle virus rL-RVG inhibits proliferation of human lung adenocarcinoma cell lines

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