冻融肿瘤疫苗联合顺铂对小鼠恶性黑色素瘤抗瘤作用的实验研究
发布时间：2018-10-14 17:40
【摘要】： 目的：探讨冻融肿瘤疫苗联合顺铂对C57BL／6J小鼠恶性黑色素瘤的抗肿瘤作用。 方法：1)将B16Fo恶性黑色素瘤细胞株接种于C57BL／6J小鼠背部皮下,建立肿瘤模型,取氩氦刀冻融后坏死肿瘤组织,联合弗氏不完全佐剂制成肿瘤疫苗。2)24只C57BL／6J小鼠随机分成4组,分别为对照组、化疗组、疫苗组及联合组(注射疫苗+顺铂),疫苗于接种瘤细胞前4周开始肌注给药,每周1次,共4次,顺铂于接种瘤细胞后3天开始腹腔注射给药,每天1次,共7次。化疗组和联合组分别于每次疫苗接种及化疗后眼眶采血,测量外周血CD4+、CD8+变化,并观察各组小鼠的出瘤情况、生存时间,测量肿瘤长径和短径变化。 结果： 1)各组小鼠成瘤率均为100%。疫苗组、联合组的平均出瘤时间分别为11.3±1.6天、11.5±1.9天,对照组、化疗组的平均出瘤时间分别为6.7±1.2天、7.7±0.8天。疫苗组、联合组分别和对照组、化疗组比较,差异均有显著性(P0.01)。 2)观察期内对照组、化疗组、疫苗组、联合组的平均生存时间分别为25.7±1.5天、37.0±2.8天、35.8±2.8天、38.5±2.1天,疫苗组、化疗组、联合组分别与对照组比较,差异均有显著性(P0.01)。 3)化疗组中,化疗2周后与化疗前比较,小鼠外周血中CD4+显著降低(P0.01),CD4+／CD8+降低(P0.05),CD8+变化无统计学差异。联合组中,第四次接种疫苗后与接种疫苗前相比,外周血中CD4+CD4+／CD8+明显升高,而CD8+下降,差异均有显著性(P0.01)；化疗后2周与接种疫苗前相比,CD8+下降、CD4+／CD8+上升,差异均有显著性(P0.05),CD4+变化无统计学差异,与第四次接种疫苗后相比,CD4+、CD4+／CD8+降低,差异均有显著性(P0.01),CD8+变化无统计学差异。 结论： 1)冻融肿瘤疫苗能有效延缓小鼠恶性黑色素瘤的生长,能延长荷瘤小鼠的生存期,改善机体的免疫状况。 2)顺铂对恶性黑色素瘤的生长有一定的抑制作用,能延长荷瘤小鼠的生存期,对荷瘤小鼠的的免疫功能有损害。 3)顺铂联合应用冻融肿瘤疫苗可弥补单纯化疗对荷瘤小鼠免疫功能的损害。
[Abstract]:Objective: to investigate the anti-tumor effect of freeze-thawed tumor vaccine combined with cisplatin on malignant melanoma in C57BL/6J mice. Methods: 1) the B16Fo malignant melanoma cell line was inoculated subcutaneously on the back of C57BL/6J mice, and the tumor model was established, and the necrotic tumor tissue was obtained after cryopreservation and thawing with argon-helium knife. 24 C57BL/6J mice were randomly divided into 4 groups: control group, chemotherapy group, vaccine group and combination group (injection of cisplatin). The vaccine was injected intramuscularly 4 weeks before inoculation of tumor cells. Cisplatin was injected intraperitoneally 3 days after inoculation of tumor cells once a week for a total of 7 times. The changes of CD4 and CD8 in peripheral blood were measured in the chemotherapy group and the combined group respectively after each inoculation and chemotherapy. The tumor emergence, survival time, tumor length and short diameter of the mice in each group were observed. Results: 1) the tumorigenesis rate of each group was 100. The mean time of tumor emergence in vaccine group and combined group was 11.3 卤1.6 days and 11.5 卤1.9 days, respectively. The average time of tumor emergence in control group and chemotherapy group was 6.7 卤1.2 days and 7.7 卤0.8 days, respectively. The average survival time of the vaccine group, the combined group and the control group were 25.7 卤1.5 days, 37.0 卤2.8 days, 35.8 卤2.8 days, 38.5 卤2.1 days respectively during the observation period (P0.01), and the average survival time of the control group, chemotherapy group, vaccine group and combined group were 25.7 卤1.5 days, 37.0 卤2.8 days, 35.8 卤2.8 days, 38.5 卤2.1 days, respectively. There were significant differences between vaccine group, chemotherapy group and combination group compared with control group (P0.01). 3) in the chemotherapy group, 2 weeks after chemotherapy and before chemotherapy, the difference was significant (P0.01). The levels of CD4 and CD4 / CD8 in peripheral blood of mice were significantly decreased (P0.01) and CD4 / CD8 were decreased (P0.05). There was no significant difference in the changes of CD8. In the combined group, the ratio of CD4 / CD8 in peripheral blood after the fourth vaccination was significantly higher than that before vaccination, but the CD8 decreased significantly (P0.01), the CD8 decreased and the CD4 / CD8 increased at 2 weeks after chemotherapy compared with that before vaccination. The difference was significant (P0.05), there was no statistical difference in CD4, compared with the fourth vaccination, CD4, CD4 / CD8 decreased, the difference was significant (P0.01), there was no significant difference in CD8. Conclusion: 1) the freeze-thaw tumor vaccine can effectively delay the growth of malignant melanoma in mice and prolong the survival time of tumor bearing mice. 2) Cisplatin can inhibit the growth of malignant melanoma and prolong the survival time of mice bearing malignant melanoma. 3) Cisplatin combined with freeze-thawed tumor vaccine can make up for the damage of immune function of tumor bearing mice caused by chemotherapy alone.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R739.5
本文编号：2271157
[Abstract]:Objective: to investigate the anti-tumor effect of freeze-thawed tumor vaccine combined with cisplatin on malignant melanoma in C57BL/6J mice. Methods: 1) the B16Fo malignant melanoma cell line was inoculated subcutaneously on the back of C57BL/6J mice, and the tumor model was established, and the necrotic tumor tissue was obtained after cryopreservation and thawing with argon-helium knife. 24 C57BL/6J mice were randomly divided into 4 groups: control group, chemotherapy group, vaccine group and combination group (injection of cisplatin). The vaccine was injected intramuscularly 4 weeks before inoculation of tumor cells. Cisplatin was injected intraperitoneally 3 days after inoculation of tumor cells once a week for a total of 7 times. The changes of CD4 and CD8 in peripheral blood were measured in the chemotherapy group and the combined group respectively after each inoculation and chemotherapy. The tumor emergence, survival time, tumor length and short diameter of the mice in each group were observed. Results: 1) the tumorigenesis rate of each group was 100. The mean time of tumor emergence in vaccine group and combined group was 11.3 卤1.6 days and 11.5 卤1.9 days, respectively. The average time of tumor emergence in control group and chemotherapy group was 6.7 卤1.2 days and 7.7 卤0.8 days, respectively. The average survival time of the vaccine group, the combined group and the control group were 25.7 卤1.5 days, 37.0 卤2.8 days, 35.8 卤2.8 days, 38.5 卤2.1 days respectively during the observation period (P0.01), and the average survival time of the control group, chemotherapy group, vaccine group and combined group were 25.7 卤1.5 days, 37.0 卤2.8 days, 35.8 卤2.8 days, 38.5 卤2.1 days, respectively. There were significant differences between vaccine group, chemotherapy group and combination group compared with control group (P0.01). 3) in the chemotherapy group, 2 weeks after chemotherapy and before chemotherapy, the difference was significant (P0.01). The levels of CD4 and CD4 / CD8 in peripheral blood of mice were significantly decreased (P0.01) and CD4 / CD8 were decreased (P0.05). There was no significant difference in the changes of CD8. In the combined group, the ratio of CD4 / CD8 in peripheral blood after the fourth vaccination was significantly higher than that before vaccination, but the CD8 decreased significantly (P0.01), the CD8 decreased and the CD4 / CD8 increased at 2 weeks after chemotherapy compared with that before vaccination. The difference was significant (P0.05), there was no statistical difference in CD4, compared with the fourth vaccination, CD4, CD4 / CD8 decreased, the difference was significant (P0.01), there was no significant difference in CD8. Conclusion: 1) the freeze-thaw tumor vaccine can effectively delay the growth of malignant melanoma in mice and prolong the survival time of tumor bearing mice. 2) Cisplatin can inhibit the growth of malignant melanoma and prolong the survival time of mice bearing malignant melanoma. 3) Cisplatin combined with freeze-thawed tumor vaccine can make up for the damage of immune function of tumor bearing mice caused by chemotherapy alone.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R739.5
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