Cell Death Mechanisms in Esophageal Squamous Cell Carcinoma Induced by Vesicular Stomatitis Virus Matrix Protein.
Cell Death Mechanisms in Esophageal Squamous Cell Carcinoma Induced by Vesicular Stomatitis Virus Matrix Protein.
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DOI：
10.24171/j.phrp.2019.10.4.08
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发表时间：
2019-08-01
影响因子：
4.3
通讯作者：
Moradi, Abdolvahab
中科院分区：
文献类型：
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作者：
Douzandegan, Yousef;Tahamtan, Alireza;Moradi, Abdolvahab
目的：水疱性口炎病毒（VSV）由于其在癌细胞中的优先复制和溶瘤活性而被开发为溶瘤病毒，然而负责的病毒成分尚未确定。本研究研究了VSV野生型（wt）和m51r突变型基质蛋白（M51R-mMP）对食管癌细胞系（KYSE-30）凋亡、焦亡、坏死和自噬途径的影响。方法：用编码wt或M51R-mMP的pcDNA3.1质粒转染KYSE-30细胞，分别在转染后48和72小时观察细胞凋亡、焦亡、坏死和自噬情况。结果：转染VSV wt和M51R-mMPs的KYSE-30细胞在转染后72小时显著降低细胞活力至< 50%。M51R-MP分别在转染后48和72 h显著提高caspase-8和caspase-9的浓度（p < 0.05）。相比之下，用VSV wt质粒转染后未检测到显著变化。此外，VSV wt和M51R-mMP转染的细胞没有改变caspase-3的表达。VSV wt和M51R-mMPs在转染后48和72小时没有mMP改变caspase-1的表达（一种焦亡标志物）。然而，转染M51R-mMP和VSV wt的细胞在感染后72小时显著增加了RIP-1（坏死下垂标志物）的表达（p < 0.05）。在转染后48小时，用VSV wt或M51R-mMPs转染也能诱导自噬的生物标志物Beclin-1。结论：本研究结果提示VSV通过不同的细胞死亡途径对KYSE-30肿瘤细胞发挥溶瘤活性，提示M51R-mMP可能具有增强溶瘤作用。
OBJECTIVES: Vesicular stomatitis virus (VSV) is under development as an oncolytic virus due to its preferential replication in cancer cells and oncolytic activity, however the viral components responsible have not yet been determined. In this study the effects of VSV wild-type (wt) and M51R-mutant matrix proteins (M51R-mMP) on apoptosis, pyroptosis, necroptosis, and autophagy pathways, in an esophagus cancer cell line (KYSE-30) were investigated.METHODS: The KYSE-30 cells were transfected with pcDNA3.1 plasmids encoding wt or M51R-mMP, and apoptosis, pyroptosis, necroptosis, and autophagy were evaluated 48 and 72 hours after transfection.RESULTS: KYSE-30 cells transfected with VSV wt and M51R-mMPs significantly reduced cell viability to < 50% at 72 hours post-transfection. M51R-MP significantly increased the concentration of caspase-8 and caspase-9 at 48 and 72 hours post-transfection, respectively ( p < 0.05). In contrast, no significant changes were detected following transfection with the VSV wt plasmid. Moreover, VSV wt and M51R-mMP transfected cells did not change the expression of caspase-3. VSV wt and M51R-mMPs did not mMP change caspase-1 expression (a marker of pyroptosis) at 48 and 72 hours post-transfection. However, M51R-mMP and VSV wt transfected cells significantly increased RIP-1 (a marker of necroptosis) expression at 72 hours post-infection ( p < 0.05). Beclin-1, a biomarker of autophagy, was also induced by transfection with VSV wt or M51R-mMPs at 48 hours post-transfection.CONCLUSION: The results in this study indicated that VSV exerts oncolytic activity in KYSE-30 tumor cells through different cell death pathways, suggesting that M51R-mMP may potentially be used to enhance oncolysis.