Effect and mechanism of the oncolytic virus Rigvir on colorectal cancer cell lines <italic style="font-style: italic">in vitro</italic>

Huang Yixin; Zhang Ruyi; Tang Jingling; Wang Xinjun

doi:10.13406/j.cnki.cyxb.003792

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Effect and mechanism of the oncolytic virus Rigvir on colorectal cancer cell lines in vitro

Precision medicine in cancer and translational studies | 更新时间：2025-10-21

- Effect and mechanism of the oncolytic virus Rigvir on colorectal cancer cell lines in vitro
- Journal of Chongqing Medical University Vol. 50, Issue 10, Pages: 1353-1362(2025)
- 作者机构：
  
  1.贵州医科大学附属医院肛肠外科，贵阳 550004
  2.贵州医科大学附属医院临床医学研究中心，贵阳 550004
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.003792
  CLC： R735.34
- Received：15 April 2024，
  
  Revised：2025-02-26，
  
  Accepted：06 March 2025，
  
  Published：28 October 2025
- 稿件说明：
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黄伊鑫,张汝一,唐景玲,等.溶瘤病毒Rigvir^®对结直肠癌细胞系在体外的影响及其机制的研究[J].重庆医科大学学报,2025,50(10):1353-1362.

Huang Yixin,Zhang Ruyi,Tang Jingling,et al.Effect and mechanism of the oncolytic virus Rigvir on colorectal cancer cell lines in vitro[J].Journal of Chongqing Medical University,2025,50(10):1353-1362.
黄伊鑫,张汝一,唐景玲,等.溶瘤病毒Rigvir^®对结直肠癌细胞系在体外的影响及其机制的研究[J].重庆医科大学学报,2025,50(10):1353-1362. DOI： 10.13406/j.cnki.cyxb.003792.

Huang Yixin,Zhang Ruyi,Tang Jingling,et al.Effect and mechanism of the oncolytic virus Rigvir on colorectal cancer cell lines in vitro[J].Journal of Chongqing Medical University,2025,50(10):1353-1362. DOI： 10.13406/j.cnki.cyxb.003792.

摘要

目的

探究溶瘤病毒Rigvir

在体外对6种不同结直肠癌细胞系的杀伤作用及不同细胞系对Rigvir

敏感性的差异，分析敏感细胞与不敏感细胞的差异基因及信号通路以探究其差异产生的原因并进行验证，根据实验结果初步探究溶瘤病毒Rigvir

对结直肠癌细胞的杀伤机制，为溶瘤病毒Rigvir

作为新型免疫治疗药物治疗结直肠癌提供理论基础。

方法

采用细胞计数试剂盒-8（cell counting kit-8，CCK-8）试验定量检测Rigvir

对细胞的细胞抑制率，通过Annexin V-FITC\PI法检测敏感细胞系的细胞凋亡率并初步分析其杀伤机制，利用生物信息学技术分析敏感细胞与不敏感细胞的差异基因及信号通路并应用蛋白质印迹法（Western blot，WB）进行验证和检测凋亡因子表达情况。高表达差异信号通路上游因子，并应用实时定量逆转录酶链式反应（real-time quantitative reverse transcription polymerase chain reaction，RT-qPCR）和WB检测Rigvir

对敏感细胞系过表达细胞株目的基因及目的蛋白表达量的影响。

结果

SW480和HT29对溶瘤病毒Rigvir

敏感，而对其余4种细胞系相对不敏感（

0.001），但2个浓度梯度的Rigvir

对HT29的抑制作用更加明显（

0.001），在感染48 h后SW480细胞随感染时间的延长，细胞抑制率不再增加（

=52.010，

=0.147），但其对病毒浓度的变化较敏感（

=13.490，

0.001）。相反，病毒浓度的变化对HT29细胞的抑制率无明显影响（

=8.450，

=0.281），但在48 h后随着感染时间的延长，抑制率仍继续升高（

=24.380，

0.001）。敏感组细胞随着感染时间的延长，凋亡率逐渐增加（

0.001），且均以细胞晚期凋亡和坏死细胞为主。通过生物信息学技术观察到PI3K/Akt/mTOR经典信号通路在敏感组与不敏感组之间差异明显，WB实验表明，应用Rigvir

后，敏感细胞组PI3K/Akt/mTOR上游蛋白表达量明显降低（

0.001）。细胞凋亡因子caspase-3及caspase-8表达量升高，Bcl-2/Bax比值降低（

0.001）。RT-qPCR和WB结果表明，Rigvir

作用于敏感细胞系HT29过表达细胞株后，PI3K/Akt/mTOR上下游信号分子（Akt，4EBP1和p70S6K）的表达量较对照组明显降低（

0.05）。

结论

Rigvir

可以在体外有效杀伤部分结直肠癌细胞系（SW480、HT29），其作用机制部分是通过PI3K/Akt/mTOR经典信号通路诱导细胞凋亡。

Abstract

Objective

To investigate the killing effect of the oncolytic virus Rigvir on six different colorectal cancer cell lines

in vitro

and differences in the sensitivity of different cell lines to Rigvir，to analyze the differentially expressed genes and signaling pathways between sensitive and insensitive cells and the reasons for such differences，to explore the killing mechanism of the oncolytic virus Rigvir on colorectal cancer cells based on experimental results，and to lay a theoretical foundation for the use of the oncolytic virus Rigvir as a novel immunotherapeutic drug for the treatment of colorectal cancer.

Methods

Cell Counting Kit-8（CCK-8） assay was used for quantitative assessment of the inhibition rate of Rigvir on cells，and the Annexin V-FITC\PI method was used to measure the apoptosis rate of sensitive cell lines and preliminarily analyze its killing mechanism. Bioinformatics techniques were used to investigate the differentially expressed genes and signaling pathways between sensitive and insensitive cells，and Western blot experiments were used for validation and detecting the expression of apoptosis factors.High expression of upstream factors in differential signaling pathways，and application of real-time quantitative reverse transcription polymerase chain reaction（RT-qPCR） and WB to detect the effect of Rigvir on the expression levels of target genes and proteins in overexpressing sensitive cell lines.

Results

SW480 and HT29 were sensitive to the oncolytic virus Rigvir，others had relatively insensitive（

0.001）. However，the inhibitory effect of Rigvir with two concentration gradients on HT29 was more significant（

0.001）. After 48 hours of infection，the cell inhibition rate of SW480 cells no longer increased with the prolongation of infection time（

52.010

，P=

0.147），but it was more sensitive to changes in virus concentration（

13.490

，P<

0.001）. On the contrary，changes in virus concentration had no significant effect on the inhibition rate of HT29 cells（

8.450

，P=

0.281），but the inhibition rate continued to increase after 48 hours with the prolongation of infection time（

24.380

，P<

0.001）. The apoptosis rate of sensitive group cells gradually increased with the prolongation of infection time（

0.001），which mainly characterized by late stage apoptosis and necrotic cells. Through bioinformatics techniques，significant differences were observed in the classic PI3K/Akt/mTOR signaling pathway between the sensitive and insensitive groups. Western blot experiments showed that after the application of Rigvir，the upstream protein expression of PI3K/Akt/mTOR in the sensitive cell group was significantly reduced（

0.001）. The expression levels of apoptosis factors caspase-3 and cas

pase-8 increased，while the Bcl-2/Bax ratio decreased（

0.001）. The results of RT-qPCR and Western blot showed that after Rigvir was applied to the sensitive cell line HT29 overexpression type，the expression levels of PI3K/Akt/mTOR upstream and downstream signaling molecules（Akt，4EBP1，and p70S6K） were significantly reduced compared to the control group（

0.05）.

Conclusion

Rigvir can effectively kill some colorectal cancer cell lines（SW480，HT29）

in vitro

，its mechanism of action is partially induced by the PI3K/Akt/mTOR classical signaling pathway to induce cell apoptosis.

关键词

Keywords

references

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⁰

Effect and mechanism of the oncolytic virus Rigvir on colorectal cancer cell lines in vitro

DOI：10.13406/j.cnki.cyxb.003792

摘要

Abstract

关键词

Keywords

references