HPV治疗性重组融合蛋白疫苗的抗肿瘤作用及机制初步探讨
发布时间：2018-10-14 16:56
【摘要】：宫颈癌是女性常见的恶性肿瘤,对于中晚期患者通常放化疗和手术治疗的效果并不理想。自从发现宫颈癌与HPV感染密切相关后,HPV治疗性疫苗成为一个重要的研究方向。本课题组前期构建了针对HPV16的mdNCRT/E7/hsp重组融合蛋白疫苗,并通过体外实验证明了该疫苗可诱导到较高水平的CTL活性及抑制血管生长作用。本研究进一步通过体内试验研究其抗肿瘤作用及其分子机制。 我们建立了C57BL/6小鼠TC-1肿瘤模型,体内实验结果表明mdNCRT/E7/hsp重组融合蛋白疫苗能够有效的抗肿瘤生长,诱发特异的肿瘤排斥反应,对未形成实体瘤的微小肿瘤病灶具有治疗作用。将两组肿瘤组织切片进行免疫组化分析,发现mdNCRT/E7/hsp重组融合蛋白能够减少肿瘤新生血管的形成,降低肿瘤的生长速度,减少发生转移浸润的机会。 将两组肿瘤细胞进行Western blot来初步研究mdNCRT/E7/hsp重组融合蛋白疫苗抗肿瘤生长的分子机制。(1)IL-6/pStat3信号传导通路：结果显示疫苗组的IL-6、p-Stat3、VEGF、HIF-1α、Snail蛋白表达水平明显下降,活化的Caspase-3蛋白水平明显提高。说明mdNCRT/E7/hsp重组融合蛋白疫苗能够通过抑制IL-6和pStat3以及其下游相关基因的表达来抑制血管生成及促进了肿瘤细胞的凋亡。(2)MAPK-ERK信号传导通路：结果显示疫苗组细胞中B-raf、p-MEK、p-ERK的表达降低,说明mdNCRT/E7/hsp重组融合蛋白疫苗能够通过抑制MAPK-ERK信号通路链中相关MAPK激酶活性,抑制肿瘤细胞的生长。 本研究结果表明：mdNCRT/E7/hsp重组融合蛋白疫苗可能通过抑制IL-6/pStat3信号传导通路和MAPK-ERK信号传导通路来抑制肿瘤新生血管的形成和肿瘤浸润转移,最终产生有效的抗肿瘤作用。
[Abstract]:Cervical cancer is a common malignant tumor in women. Since the discovery of close association between cervical cancer and HPV infection, HPV therapeutic vaccine has become an important research direction. The recombinant mdNCRT/E7/hsp fusion protein vaccine against HPV16 was constructed in our research group, and it was proved in vitro that the vaccine could induce higher CTL activity and inhibit angiogenesis. In this study, the anti-tumor effect and its molecular mechanism were studied in vivo. The TC-1 tumor model of C57BL/6 mice was established. The results of in vivo experiments showed that the recombinant mdNCRT/E7/hsp fusion protein vaccine could effectively inhibit tumor growth, induce specific tumor rejection, and have therapeutic effect on small tumor lesions without solid tumors. Immunohistochemical analysis of the two groups showed that the recombinant mdNCRT/E7/hsp fusion protein could reduce the angiogenesis of tumor, decrease the growth rate of tumor and reduce the chance of metastasis and infiltration. Two groups of tumor cells were treated with Western blot to study the molecular mechanism of mdNCRT/E7/hsp recombinant fusion protein vaccine against tumor growth. (1) IL-6/pStat3 signal transduction pathway: the results showed that the expression of IL-6,p-Stat3,VEGF,HIF-1 伪 and Snail protein decreased significantly and the activated Caspase-3 protein level increased significantly in the vaccine group. The results showed that mdNCRT/E7/hsp recombinant fusion protein vaccine could inhibit angiogenesis and promote apoptosis of tumor cells by inhibiting the expression of IL-6, pStat3 and its downstream genes. (2) MAPK-ERK signaling pathway: the results showed that the expression of B-raftip-MEKp-ERK was decreased in the cells of vaccine group. The results showed that the recombinant mdNCRT/E7/hsp fusion protein vaccine could inhibit the growth of tumor cells by inhibiting the activity of MAPK kinase in the MAPK-ERK signaling pathway chain. Our results suggest that the recombinant mdNCRT/E7/hsp fusion protein vaccine may inhibit angiogenesis, tumor invasion and metastasis by inhibiting IL-6/pStat3 signal transduction pathway and MAPK-ERK signal transduction pathway, resulting in an effective anti-tumor effect.
【学位授予单位】：北京协和医学院
【学位级别】：博士
【学位授予年份】：2011
【分类号】：R392
本文编号：2271051
[Abstract]:Cervical cancer is a common malignant tumor in women. Since the discovery of close association between cervical cancer and HPV infection, HPV therapeutic vaccine has become an important research direction. The recombinant mdNCRT/E7/hsp fusion protein vaccine against HPV16 was constructed in our research group, and it was proved in vitro that the vaccine could induce higher CTL activity and inhibit angiogenesis. In this study, the anti-tumor effect and its molecular mechanism were studied in vivo. The TC-1 tumor model of C57BL/6 mice was established. The results of in vivo experiments showed that the recombinant mdNCRT/E7/hsp fusion protein vaccine could effectively inhibit tumor growth, induce specific tumor rejection, and have therapeutic effect on small tumor lesions without solid tumors. Immunohistochemical analysis of the two groups showed that the recombinant mdNCRT/E7/hsp fusion protein could reduce the angiogenesis of tumor, decrease the growth rate of tumor and reduce the chance of metastasis and infiltration. Two groups of tumor cells were treated with Western blot to study the molecular mechanism of mdNCRT/E7/hsp recombinant fusion protein vaccine against tumor growth. (1) IL-6/pStat3 signal transduction pathway: the results showed that the expression of IL-6,p-Stat3,VEGF,HIF-1 伪 and Snail protein decreased significantly and the activated Caspase-3 protein level increased significantly in the vaccine group. The results showed that mdNCRT/E7/hsp recombinant fusion protein vaccine could inhibit angiogenesis and promote apoptosis of tumor cells by inhibiting the expression of IL-6, pStat3 and its downstream genes. (2) MAPK-ERK signaling pathway: the results showed that the expression of B-raftip-MEKp-ERK was decreased in the cells of vaccine group. The results showed that the recombinant mdNCRT/E7/hsp fusion protein vaccine could inhibit the growth of tumor cells by inhibiting the activity of MAPK kinase in the MAPK-ERK signaling pathway chain. Our results suggest that the recombinant mdNCRT/E7/hsp fusion protein vaccine may inhibit angiogenesis, tumor invasion and metastasis by inhibiting IL-6/pStat3 signal transduction pathway and MAPK-ERK signal transduction pathway, resulting in an effective anti-tumor effect.
【学位授予单位】：北京协和医学院
【学位级别】：博士
【学位授予年份】：2011
【分类号】：R392
【参考文献】
相关期刊论文 前10条
1 吴民华;陈小毅;;STAT3表达与肿瘤关系的研究进展[J];广东医学院学报;2007年05期
2 刘涛,田聆,魏于全,赵菊梅,梁传余;热休克蛋白与肿瘤的研究进展[J];华西医学;2004年03期
3 林晓华;吴宁;侯丽辉;吴效科;;人乳头瘤病毒与宫颈肿瘤关系的研究进展[J];医学研究生学报;2006年02期
4 赵立民;罗红敏;王春英;李翎;;rBCG-HPV16 E6-E7疫苗的构建和免疫原性研究[J];中华肿瘤防治杂志;2006年08期
5 刘丽丽;战雪梅;孙崇伟;孙莉莉;;转录信号传导子与激活子3在宫颈癌组织中的表达及临床意义[J];中华肿瘤防治杂志;2007年06期
6 许雪梅;;人乳头瘤病毒及宫颈癌疫苗的研究——解读2008年诺贝尔生理学或医学奖[J];生物化学与生物物理进展;2008年10期
7 周慧,李盛贤,平文祥;佐剂的研究进展[J];生物技术;2004年04期
8 蒋业贵;王宇明;李奇芬;;HLA-DR抗原和HSP70在肝细胞癌中的表达意义[J];世界华人消化杂志;2001年10期
9 谢晨曦;任建林;;P38 MAPK信号传导通路与胃癌关系的研究现状[J];世界华人消化杂志;2008年30期
10 徐道华;周晨慧;夏永鹏;邱宗荫;贾正才;周伟;;宫颈癌治疗性多肽疫苗的合成及其诱导细胞毒性T淋巴细胞效应的研究[J];中国临床药理学与治疗学;2007年02期
，本文编号：2271051
本文链接：https://www.wllwen.com/xiyixuelunwen/2271051.html
最近更新
教材专著
