【摘要】： 目的：探討凍融腫瘤疫苗聯(lián)合順鉑對C57BL／6J小鼠惡性黑色素瘤的抗腫瘤作用。 方法：1)將B16Fo惡性黑色素瘤細胞株接種于C57BL／6J小鼠背部皮下,建立腫瘤模型,取氬氦刀凍融后壞死腫瘤組織,聯(lián)合弗氏不完全佐劑制成腫瘤疫苗。2)24只C57BL／6J小鼠隨機分成4組,分別為對照組、化療組、疫苗組及聯(lián)合組(注射疫苗+順鉑),疫苗于接種瘤細胞前4周開始肌注給藥,每周1次,共4次,順鉑于接種瘤細胞后3天開始腹腔注射給藥,每天1次,共7次。化療組和聯(lián)合組分別于每次疫苗接種及化療后眼眶采血,測量外周血CD4+、CD8+變化,并觀察各組小鼠的出瘤情況、生存時間,測量腫瘤長徑和短徑變化。 結(jié)果： 1)各組小鼠成瘤率均為100%。疫苗組、聯(lián)合組的平均出瘤時間分別為11.3±1.6天、11.5±1.9天,對照組、化療組的平均出瘤時間分別為6.7±1.2天、7.7±0.8天。疫苗組、聯(lián)合組分別和對照組、化療組比較,差異均有顯著性(P0.01)。 2)觀察期內(nèi)對照組、化療組、疫苗組、聯(lián)合組的平均生存時間分別為25.7±1.5天、37.0±2.8天、35.8±2.8天、38.5±2.1天,疫苗組、化療組、聯(lián)合組分別與對照組比較,差異均有顯著性(P0.01)。 3)化療組中,化療2周后與化療前比較,小鼠外周血中CD4+顯著降低(P0.01),CD4+／CD8+降低(P0.05),CD8+變化無統(tǒng)計學差異。聯(lián)合組中,第四次接種疫苗后與接種疫苗前相比,外周血中CD4+CD4+／CD8+明顯升高,而CD8+下降,差異均有顯著性(P0.01)；化療后2周與接種疫苗前相比,CD8+下降、CD4+／CD8+上升,差異均有顯著性(P0.05),CD4+變化無統(tǒng)計學差異,與第四次接種疫苗后相比,CD4+、CD4+／CD8+降低,差異均有顯著性(P0.01),CD8+變化無統(tǒng)計學差異。 結(jié)論： 1)凍融腫瘤疫苗能有效延緩小鼠惡性黑色素瘤的生長,能延長荷瘤小鼠的生存期,改善機體的免疫狀況。 2)順鉑對惡性黑色素瘤的生長有一定的抑制作用,能延長荷瘤小鼠的生存期,對荷瘤小鼠的的免疫功能有損害。 3)順鉑聯(lián)合應用凍融腫瘤疫苗可彌補單純化療對荷瘤小鼠免疫功能的損害。
[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 鹵1.6 days and 11.5 鹵1.9 days, respectively. The average time of tumor emergence in control group and chemotherapy group was 6.7 鹵1.2 days and 7.7 鹵0.8 days, respectively. The average survival time of the vaccine group, the combined group and the control group were 25.7 鹵1.5 days, 37.0 鹵2.8 days, 35.8 鹵2.8 days, 38.5 鹵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 鹵1.5 days, 37.0 鹵2.8 days, 35.8 鹵2.8 days, 38.5 鹵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.
【學位授予單位】：中南大學
【學位級別】：碩士
【學位授予年份】：2010
【分類號】：R739.5

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