猪A型塞内卡毒株CH-01-2015的溶瘤效果

Oncolytic Effect of Swine Senecavirus A Strain CH-01-2015

为探究猪A型塞内卡毒株(SVA)CH-01-2015的溶瘤效果,本试验以SVA CH-01-2015为研究对象,在体外通过显微镜观察和细胞活性测定检测该毒株对前列腺癌细胞(PC-3)、非小细胞肺癌细胞(H1299、A549)、子宫内膜癌细胞(Ishikawa)、胶质瘤细胞(U251)和宫颈癌细胞(Hela)共6种肿瘤细胞的杀伤情况,通过病毒复制动力学测定检测SVA CH-01-2015在不同肿瘤细胞中的复制情况;在体内,通过PC-3裸鼠荷瘤试验检测SVA CH-01-2015的溶瘤效果,通过免疫组化和组织病理学染色检测肿瘤组织中病毒的复制以及治疗后肿瘤组织的形态变化。体外试验结果显示,与对照组细胞相比,感染SVA CH-01-2015的PC-3、H1299和Ishikawa肿瘤细胞被显著裂解,而U251、A549和Hela肿瘤细胞形态无显著变化;且与对照组细胞相比,SVA CH-01-2015毒株能极显著杀伤PC-3、H1299和Ishikawa肿瘤细胞(P<0.01),对U251、A549和Hela肿瘤细胞几乎无杀伤作用(P>0.05);以感染复数(MOI)为1的SVA CH-01-2015分别感染6种肿瘤细胞,其在PC-3、H1299和Ishikawa中复制能力极强,在U251和A549中复制能力较弱,在Hela中基本不复制。体内试验结果显示,与对照组相比,静脉单针、瘤内单针和瘤内2针注射10^(8) TCID_(50)/针的SVA CH-01-2015均能极显著抑制PC-3裸鼠移植瘤的生长(P<0.01),且瘤内2针注射的抑制效果更为显著(P<0.01)。结果表明,SVA CH-01-2015能显著杀伤PC-3、H1299和Ishikawa肿瘤细胞,对U251、A549和Hela肿瘤细胞无杀伤作用,且该病毒能抑制体内PC-3裸鼠移植瘤的生长,瘤内2针注射的抑制效果更为显著。

In order to clarify the oncolytic effect of swine Senecavirus A(SVA)strain CH-01-2015,this study examined the in vitro oncolytic effect of SVA CH-01-2015 on prostate cancer cells(PC-3),non-small cell lung cancer cells(H1299,A549),endometrial cancer cells(Ishikawa),glioma cells(U251),and cervical cancer cells(Hela)by microscope observation and cell viability assay;the replication of SVA CH-01-2015 in different tumor cells was detected by virus replication kinetics.The in vivo oncolytic effect of SVA CH-01-2015 was detected by PC-3 nude mouse tumor-bearing experiment,the virus replication in tumor tissue and the morphological changes of tumor tissue after treatment were analyzed by immunohistochemistry and histopathological staining,respectively.The results of in vitro experiments showed that compared with the control group cells,PC-3,H1299 and Ishikawa tumor cells infected with SVA CH-01-2015 were significantly lysed,while the morphology of U251,A549 and Hela tumor cells infected with the virus did not change significantly.Compared with the control group,SVA CH-01-2015 strain had significant killing effect on PC-3,H1299 and Ishikawa tumor cells(P<0.01),but almost no killing effect on U251,A549 and Hela tumor cells(P>0.05).When SVA CH-01-2015 with multiplicity of infection(MOI)of 1 was used to inoculate PC-3,H1299,Ishikawa,U251,A549 and Hela tumor cells,the growth kinetics results showed that SVA CH-01-2015 replicated efficiently on PC-3,H1299,and Ishikawa tumor cells,had a weak replication on U251 and A549 tumor cells,but it did not replicate on Hela tumor cells.The results of in vivo experiments showed that compared with the control group,intravenous single-needle injection,intertumoral single-needle injection and intertumoral double-needle injection of SVA CH-01-2015 of 10^(8) TCID_(50)/needle inhibited the growth of PC-3 xenografts in nude mice(P<0.01);and intertumoral double-needle injection inhibited the growth tumor more significantly than other injection methods(P<0.01).Thus,SVA CH-01-2015 has significant killing effect on PC-3,H1299,Ishikawa tumor cells,but has no killing effect on U251,A549 and Hela tumor cells.The virus inhibits the growth of PC-3 transplanted tumors in nude mice in vivo,and intertumoral double-needle injection of SVA CH-01-2015 produces strong oncolytic effect.