白细胞介素-24蛋白及基因联合顺铂对卵巢癌SKOV3细胞体外生长的影响

发布时间：2018-02-01 07:38

本文关键词： 白细胞介素-24 卵巢癌细胞生长抑制细胞凋亡细胞周期多基因遗传表达分析　出处：《遵义医学院》2014年硕士论文　论文类型：学位论文

【摘要】：目的： ①探讨rhIL-24蛋白与顺铂联合体外对人卵巢癌SKOV3细胞、人卵巢癌耐顺铂株SKOV3/ddp细胞的抑制效应； ②探讨rhIL-24蛋白联合顺铂体外诱导人卵巢癌SKOV3细胞、人卵巢癌耐顺铂株SKOV3/ddp细胞凋亡效应、细胞周期及凋亡基因的变化； ③探讨腺病毒介导的白细胞介素-24基因与顺铂联合体外诱导人卵巢癌SKOV3细胞的抑制效应及细胞凋亡情况变化。方法： ①用rhIL-24、DDP及rhIL-24+DDP作用于SKOV3细胞、SKOV3/ddp细胞，用Ad-hIL-24、DDP及Ad-hIL-24+DDP作用于SKOV3细胞抑制细胞生长； ②应用CCK-8法检测SKOV3细胞、SKOV3/ddp细胞增殖情况，计算抑制率； ③应用流式细胞仪检测SKOV3细胞、SKOV3/ddp细胞凋亡及细胞周期的变化； ④应用多基因遗传表达分析SKOV3细胞、SKOV3/ddp细胞凋亡多基因的变化。结果： ①SKOV3细胞、SKOV3/ddp细胞在适宜条件下培养于第3天进入对数生长期； ②rhIL-24蛋白能抑制SKOV3细胞、SKOV3/ddp细胞生长，48小时的抑制率分别为30.7%、21.8%，对正常细胞无毒性作用；联合DDP组抑制率分别为35.3%、32.6%，优于单独用药组，，rhIL-24蛋白具有化疗增敏效应； ③流式细胞仪检测rhIL-24、DDP和rhIL-24+DDP各组凋亡率分别与阴性对照组比较，差异有显著性意义；细胞周期检测中，rhIL-24作用SKOV3细胞、SKOV3/ddp细胞，分别引起G2期阻滞，G2期及S期阻滞。 ④rhIL-24蛋白对于SKOV3细胞、SKOV3/ddp细胞具有上调Bax，下调Bcl-2，下调survivin，激活caspase3,上调抑癌基因Rb等凋亡相关基因，联合DDP组中作用效果更加显著。 ⑤Ad-hIL-24能抑制SKOV3细胞的生长，且联合用药之后对细胞生长抑制率明显优于单独用药组，Ad-hIL-24具有化疗增敏效应。结论： ①rhIL-24蛋白及Ad-hIL-24均能抑制SKOV3细胞、SKOV3/ddp细胞的体外生长，分别联合DDP后效果优于单独用药组，具有化疗增敏效应，同时能诱导SKOV3细胞、SKOV3/ddp细胞凋亡。 ②rhIL-24蛋白可影响SKOV3细胞、SKOV3/ddp细胞的细胞周期，且能引起细胞凋亡相关基因的变化。
[Abstract]:Objective: 1 to investigate the inhibitory effect of rhIL-24 protein combined with cisplatin on human ovarian cancer SKOV3 cells and cisplatin resistant SKOV3/ddp cells. (2) to investigate the effect of rhIL-24 protein combined with cisplatin on inducing apoptosis of human ovarian cancer SKOV3 cells in vitro, and the changes of cell cycle and apoptotic genes in cisplatin-resistant SKOV3/ddp cells. 3 to investigate the inhibitory effect of adenovirus-mediated interleukin-24 gene and cisplatin combined with cisplatin on human ovarian cancer SKOV3 cells and the changes of apoptosis. Methods: 1 rhIL-24 DDP and rhIL-24 DDP were used to treat SKOV3 cells with SKOV3 / DDP cells and Ad-hIL-24. DDP and Ad-hIL-24 DDP inhibited the growth of SKOV3 cells. (2) the proliferation of SKOV3 / DDP cells in SKOV3 cells was detected by CCK-8 assay, and the inhibitory rate was calculated. 3The apoptosis and cell cycle of SKOV3 cell line SKOV3 / DDP were detected by flow cytometry. 4 Polygene expression was used to analyze the change of apoptotic polygene in SKOV3 cell line SKOV3 / DDP. Results: (1) SKOV3 cells entered logarithmic growth stage on the 3rd day. 2the inhibitory rates of rhIL-24 protein on the growth of SKOV3 / DDP cells in SKOV3 cells were 30.7 and 21.8h, respectively, and there was no toxic effect on normal cells. The inhibitory rates of rhIL-24 protein in combination with DDP group were 35.3and 32.6, respectively, which were superior to those of rhIL-24 protein alone in chemosensitization. 3The apoptotic rate of DDP and rhIL-24 DDP was detected by flow cytometry, which was significantly higher than that of negative control group. RhIL-24 induced the arrest of SKOV3 / DDP cells in SKOV3 cells in G2 phase and S-phase arrest, respectively. 4 rhIL-24 protein up-regulated Bax-2, down-regulated Bcl-2, down-regulated survivin and activated caspase3 in SKOV3 cell line SKOV3 / DDP. Apoptosis related genes such as RB were upregulated in combination with DDP. 5 Ad-hIL-24 could inhibit the growth of SKOV3 cells, and the inhibitory rate of Ad-hIL-24 was significantly higher than that of Ad-hIL-24 alone. Conclusion: 1rhIL-24 protein and Ad-hIL-24 could inhibit the growth of SKOV3 / DDP cells in SKOV3 cells in vitro, and the effect after combined with DDP was better than that in the control group. It has chemosensitization effect and induces apoptosis of SKOV3 / DDP cells in SKOV3 cells. 2rhIL-24 protein can affect the cell cycle of SKOV3 / DDP cells in SKOV3 cells and induce the changes of apoptosis-related genes.
【学位授予单位】：遵义医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.31

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