IL-24联合顺铂对人宫颈癌裸鼠移植瘤抑制及化疗增敏作用的探究
发布时间：2018-09-19 09:51
【摘要】：目的 通过肿瘤动物模型,探讨基因重组质粒白细胞介素-24(pDC316-hIL-24).顺铂(DDP)单独及联合干预对人宫颈癌Siha细胞株裸鼠移植瘤抑制及其化疗增敏作用的探究,为宫颈癌临床治疗提供一定的理论基础。 方法 将人宫颈癌Siha细胞株接种BALB/c雌性裸鼠,构建人宫颈癌裸鼠移植瘤动物模型,随机将符合成瘤标准的动物模型分为6组,每组6只,分别是：PBS缓冲液对照组(PBS组)、空质粒pDC316对照组(空质粒组)、基因重组质粒pDC316-hIL-24组(IL-24组)、半量顺铂组(半DDP组)、全量顺铂组(全DDP组)、IL-24基因重组质粒+半量顺铂组(联合组)。观察及比较干预后裸鼠的一般情况、体重改变及肿瘤生长状况；实验结束后摘取眼球取血,脱臼法处死裸鼠,剥取瘤体、留取脾脏,分别计算抑瘤率和脾指数；应用聚合酶链反应(PCR)法鉴定移植瘤中IL-24是否成功转染及表达；酶联免疫吸附测定(ELISA)法检测裸鼠血肿瘤坏死因子α(TNF-α)和γ-干扰素(IFN-γ)浓度；蛋白印记(Western blot)法观察移植瘤增殖细胞核抗原(PCNA)的表达；免疫组化染色法观察移植瘤碱性成纤维细胞生长因子(b-FGF)及微血管密度(MVD)的变化。 结果 1.IL-24基因重组质粒在移植瘤中被成功转染及表达。2.联合组、IL-24组、全量DDP组抑瘤率分别为51.5%、33.9%、36.3%高于PBS对照组、空质粒组、半量DDP组(P0.001),其中联合组抑瘤率明显优于其他组(P0.001),且联合组、IL-24组裸鼠饮食、活动、精神状态较好,IL-24组裸鼠体重较对照组无明显改变(P0.05),PBS组和空质粒组抑瘤率差异无统计学意义(P0.05)。3.联合组、IL-24组血清TNF-α、IFN-γ、脾指数与其他组比较升高有统计学意义(P0.001),全量顺铂组血清TNF-α、IFN-γ、脾指数与其他组比较降低有统计学意义(P0.001)。4.Western blot、免疫组化结果显示联合组、IL-24组移植瘤中PCNA、 b-FGF、MVD表达下调较对照组比较有统计学意义(P0.01),其中联合干预组下调的幅度更大。 结论 1.IL-24、顺铂联合应用对裸鼠宫颈癌移植瘤生长有明显的抑制作用,且效果强于两者单独作用。2.IL-24可增强宫颈癌对化疗药物顺铂的敏感性并减少其单纯化疗的毒副作用。3.这些作用可能与IL-24通过调节机体免疫功能,抑制肿瘤的增殖和微血管形成有关。
[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 伪 (TNF- 伪) and interferon 纬 (IFN- 纬) 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- 伪 -IFN- 纬 and spleen index of the combined group were significantly higher than those of the other groups (P0.001), while the serum TNF- 伪 -IFN- 纬 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.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.33
本文编号：2249785
[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 伪 (TNF- 伪) and interferon 纬 (IFN- 纬) 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- 伪 -IFN- 纬 and spleen index of the combined group were significantly higher than those of the other groups (P0.001), while the serum TNF- 伪 -IFN- 纬 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.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.33
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