【摘要】：目的 通過腫瘤動物模型,探討基因重組質(zhì)粒白細胞介素-24(pDC316-hIL-24).順鉑(DDP)單獨及聯(lián)合干預對人宮頸癌Siha細胞株裸鼠移植瘤抑制及其化療增敏作用的探究,為宮頸癌臨床治療提供一定的理論基礎。 方法 將人宮頸癌Siha細胞株接種BALB/c雌性裸鼠,構建人宮頸癌裸鼠移植瘤動物模型,隨機將符合成瘤標準的動物模型分為6組,每組6只,分別是：PBS緩沖液對照組(PBS組)、空質(zhì)粒pDC316對照組(空質(zhì)粒組)、基因重組質(zhì)粒pDC316-hIL-24組(IL-24組)、半量順鉑組(半DDP組)、全量順鉑組(全DDP組)、IL-24基因重組質(zhì)粒+半量順鉑組(聯(lián)合組)。觀察及比較干預后裸鼠的一般情況、體重改變及腫瘤生長狀況；實驗結束后摘取眼球取血,脫臼法處死裸鼠,剝?nèi)×鲶w、留取脾臟,分別計算抑瘤率和脾指數(shù)；應用聚合酶鏈反應(PCR)法鑒定移植瘤中IL-24是否成功轉染及表達；酶聯(lián)免疫吸附測定(ELISA)法檢測裸鼠血腫瘤壞死因子α(TNF-α)和γ-干擾素(IFN-γ)濃度；蛋白印記(Western blot)法觀察移植瘤增殖細胞核抗原(PCNA)的表達；免疫組化染色法觀察移植瘤堿性成纖維細胞生長因子(b-FGF)及微血管密度(MVD)的變化。 結果 1.IL-24基因重組質(zhì)粒在移植瘤中被成功轉染及表達。2.聯(lián)合組、IL-24組、全量DDP組抑瘤率分別為51.5%、33.9%、36.3%高于PBS對照組、空質(zhì)粒組、半量DDP組(P0.001),其中聯(lián)合組抑瘤率明顯優(yōu)于其他組(P0.001),且聯(lián)合組、IL-24組裸鼠飲食、活動、精神狀態(tài)較好,IL-24組裸鼠體重較對照組無明顯改變(P0.05),PBS組和空質(zhì)粒組抑瘤率差異無統(tǒng)計學意義(P0.05)。3.聯(lián)合組、IL-24組血清TNF-α、IFN-γ、脾指數(shù)與其他組比較升高有統(tǒng)計學意義(P0.001),全量順鉑組血清TNF-α、IFN-γ、脾指數(shù)與其他組比較降低有統(tǒng)計學意義(P0.001)。4.Western blot、免疫組化結果顯示聯(lián)合組、IL-24組移植瘤中PCNA、 b-FGF、MVD表達下調(diào)較對照組比較有統(tǒng)計學意義(P0.01),其中聯(lián)合干預組下調(diào)的幅度更大。 結論 1.IL-24、順鉑聯(lián)合應用對裸鼠宮頸癌移植瘤生長有明顯的抑制作用,且效果強于兩者單獨作用。2.IL-24可增強宮頸癌對化療藥物順鉑的敏感性并減少其單純化療的毒副作用。3.這些作用可能與IL-24通過調(diào)節(jié)機體免疫功能,抑制腫瘤的增殖和微血管形成有關。
[Abstract]:Objective to study the recombinant plasmid interleukin-24 (pDC316-hIL-24) in animal model of tumor. The study of the inhibitory effect of cisplatin (DDP) alone or in combination on human cervical carcinoma Siha cell line in nude mice and its chemotherapeutic sensitizing effect provides a theoretical basis for clinical treatment of cervical cancer. Methods Human cervical cancer Siha cell line was inoculated into BALB/c female nude mice to construct the transplanted tumor animal model of human cervical carcinoma. The animal models which met the criteria of tumorigenesis were randomly divided into 6 groups, 6 in each group. They were: control group (PBS group), blank plasmid pDC316 group (empty plasmid group), recombinant plasmid pDC316-hIL-24 group (IL-24 group), half-dose cisplatin group (semi-DDP group), total cisplatin group (whole DDP group), recombinant plasmid half-dose cisplatin group (combination group). The general situation, weight change and tumor growth of nude mice after intervention were observed and compared. After the experiment, the eyeball was taken and blood was taken out, the nude mice were killed by dislocated method, the tumor body was stripped, the spleen was retained, and the tumor inhibition rate and spleen index were calculated respectively. Polymerase chain reaction (PCR) was used to identify the successful transfection and expression of IL-24 in xenografts, and the concentrations of tumor necrosis factor 偽 (TNF- 偽) and interferon 緯 (IFN- 緯) in nude mice were detected by (ELISA) assay. The expression of proliferating cell nuclear antigen (PCNA) was detected by (Western blot) and the changes of basic fibroblast growth factor (b-FGF) and microvessel density (MVD) were observed by immunohistochemical staining. Results the recombinant plasmid of 1.IL-24 gene was successfully transfected and expressed in transplanted tumor. The inhibition rate of IL-24 in the combined group was 51.5% and 36.3% higher than that in the PBS control group. The inhibition rate of the combined group was significantly higher than that of the other groups (P0.001), and the inhibitory rate of the combined group was significantly higher than that of the other groups (P0.001), and the inhibitory rate of the combined group was significantly higher than that of the other groups (P0.001), and the inhibitory rate of the combined group was significantly higher than that of the control group (P0.001). The weight of Nude mice in IL-24 group had no significant change compared with the control group (P0.05). There was no significant difference in tumor inhibition rate between PBS group and empty plasmid group (P0.05). The serum TNF- 偽 -IFN- 緯 and spleen index of the combined group were significantly higher than those of the other groups (P0.001), while the serum TNF- 偽 -IFN- 緯 and the spleen index of the Cisplatin group were significantly lower than those of the other groups (P0.001). 4. The immunohistochemical results showed that the spleen index of the combined group was significantly lower than that of the other groups. 4. The immunohistochemical results showed that the spleen index of the combined group was significantly higher than that of the other groups. The down-regulation of PCNA, b-FGFN MVD was significantly lower than that of the control group (P0.01), and the down-regulation was more significant in the combined intervention group. Conclusion 1. The combination of IL-24 and cisplatin can significantly inhibit the growth of cervical cancer transplanted tumor in nude mice, and the effect is stronger than that of both alone. 2. IL-24 can enhance the sensitivity of cervical cancer to cisplatin and reduce the toxicity and side effect of chemotherapy alone. These effects may be related to the inhibition of tumor proliferation and microangiogenesis by IL-24 regulating immune function.
【學位授予單位】：山西醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R737.33

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