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中华腔镜泌尿外科杂志(电子版) ›› 2023, Vol. 17 ›› Issue (04) : 388 -393. doi: 10.3877/cma.j.issn.1674-3253.2023.04.015

实验研究

新型溶瘤病毒M1激活内质网应激致前列腺癌细胞凋亡的机制
刘硕儒, 王功炜, 张斌, 李书豪, 胡成()   
  1. 510630 广州,中山大学附属第三医院泌尿外科
  • 收稿日期:2023-03-15 出版日期:2023-08-01
  • 通信作者: 胡成
  • 基金资助:
    广东省自然科学基金(2021A1515012322)

Oncolytic virus M1 activates endoplasmic reticulum stress pathway to induce apoptosis of prostate cancer cells

Shuoru Liu, Gongwei Wang, Bin Zhang, Shuhao Li, Cheng Hu()   

  1. Department of Urology, the Third Affiliated Hospital of Sun Yat-sen University, Guangzhou 510630, China
  • Received:2023-03-15 Published:2023-08-01
  • Corresponding author: Cheng Hu
引用本文:

刘硕儒, 王功炜, 张斌, 李书豪, 胡成. 新型溶瘤病毒M1激活内质网应激致前列腺癌细胞凋亡的机制[J]. 中华腔镜泌尿外科杂志(电子版), 2023, 17(04): 388-393.

Shuoru Liu, Gongwei Wang, Bin Zhang, Shuhao Li, Cheng Hu. Oncolytic virus M1 activates endoplasmic reticulum stress pathway to induce apoptosis of prostate cancer cells[J]. Chinese Journal of Endourology(Electronic Edition), 2023, 17(04): 388-393.

目的

探讨新型溶瘤病毒M1对前列腺癌细胞的杀伤作用及其所引起凋亡的机理。

方法

通过四甲基偶氮唑盐比色法(MTT)检测不同滴度(0、0.001、0.01、0.1、1、10 PFU/cell)M1病毒对前列腺癌细胞进行48 h处理后细胞的存活率,通过流式细胞术检测前列腺癌细胞凋亡的情况,通过蛋白标记(Western Blot)检测细胞内质网应激通路和凋亡通路的关键分子。

结果

与对照组(0 PFU/cell)相比,不同滴度(0.001、0.01、0.1、1、10 PFU/cell)溶瘤病毒M1能显著降低前列腺癌细胞存活率(P<0.05),且随着M1病毒剂量增加,前列腺癌细胞凋亡率显著上升(P<0.05)。蛋白标记(Western Blot)结果显示前列腺癌感染M1病毒后,内质网应激通路CHOP、Caspase 12、凋亡通路Cleaved-caspase 3蛋白显著上调(P<0.05)。

结论

新型溶瘤病毒M1可能通过引起前列腺癌细胞发生内质网应激而产生凋亡,最终杀伤前列腺癌细胞。

Objective

To investigate if M1 virus causes apoptosis of prostate cancer cells and its mechanism.

Methods

MTT assay was used to detect survival rate of prostate cancer cells that were treated with a range of titers (0, 0.001, 0.01, 0.1, 1, 10 PFU/cell) for 48 h. Flow cytometry was used to detect the effect of different titers (0, 0.001, 0.01, 0.1 PFU/cell) on cell apoptosis after 48 h. Western Blot was performed to explore the effect of various titers (0, 0.001, 0.01, 0.1 PFU/cell) on endoplasmic reticulum stress and apoptosis pathway after 48 h.

Results

Compared with the control group (0 PFU/cell), different titers (0.001, 0.01, 0.1 PFU/cell) of M1 virus could significantly reduce the survival rate of prostate cancer cells (P<0.05). As the M1 virus dose increased, the cell apoptosis rate obviously increased (P<0.05). Western Blot displayed the level of CHOP, Caspase 12 and Cleaved-caspase 3 were significantly up-regulated after M1 infection (P<0.05).

Conclusions

Oncolytic virus M1 may induce cell apoptosis via endoplasmic reticulum stress in prostate cancer cells.

图1 溶瘤病毒M1对前列腺癌细胞的杀伤注:实验运用人前列腺癌低转移细胞PC-3M-2B4,人前列腺癌转移性细胞22RV1、PC3;a图为溶瘤病毒M1对不同前列腺癌细胞以及正常前列腺上皮细胞的杀伤,b图为PC-3M-2B4、22RV1、RWPE1分别经过溶瘤病毒M1(10 MOI)和多西他赛(12 nM)处理48 h后的细胞存活率图;图上各点均为与对照组(0 PFU/cell)相比较,**P<0.01,***P<0.001,****P<0.0001
图3 溶瘤病毒M1对内质网应激以及凋亡蛋白水平的影响
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