IP6对卵巢癌SKOV3细胞株增殖凋亡的影响

发布时间：2018-01-16 10:21

本文关键词：IP6对卵巢癌SKOV3细胞株增殖凋亡的影响　出处：《河北医科大学》2017年硕士论文　论文类型：学位论文

更多相关文章： 卵巢癌 SKOV3 IP6 Rb CyclinD1

【摘要】：卵巢癌是妇科常见的恶性肿瘤之一,其死亡率居妇科肿瘤的首位,虽然国内外有关卵巢癌的研究很多,但它的发病原因和发病机制尚未完全研究清楚。因卵巢癌早期症状不易察觉,晚期病例也缺乏有效的治疗手段,多数患者确诊时已出现腹水及远处转移等晚期症状,虽然通过手术和化疗能够明显的改善晚期卵巢癌患者的生活质量,但2017年美国国家癌症研究所调查:卵巢癌的五年生存率为45.6%[1],长期生存率更低。现如今,卵巢癌已成为严重威胁广大女性生命健康的最主要的妇科肿瘤。2012年Nature的科学家发现,一些卵巢上皮癌起源于两种上皮组织的过渡带,另一些起源于上皮组织的干细胞[2],因此,各国学者正在积极寻找上皮性卵巢癌真正的起源。研究卵巢上皮性癌发生和发展的相关机制及探求有效的分子标记物和治疗靶点[3~4]仍是目前卵巢癌研究的热点和难点。IP6(Inositol hexaphosphate)--六磷酸肌醇,是一种天然的含磷化合物,其分子结构包括肌醇和6个磷酸离子。它以植酸(肌醇六磷酸)钙镁盐的形式丰富存在于酵母、放线菌、哺乳动物和植物[5],其抗癌作用已在体内外实验中得到证实,但作用机制尚不完全清楚。IP6已成为近年来抗癌药物研究的热点,并在结肠癌,肺癌,乳腺癌,前列腺癌等多种类型的恶性肿瘤体内体外实验研究中也证明其有效的抗癌作用。目前对IP6主要的研究内容包括:对肿瘤细胞的免疫杀伤作用、促进癌细胞凋亡作用、抗突变及DNA修复作用及端粒酶的抑制作用及抗氧化作用等[6-7]。Rb基因是一种肿瘤抑制基因,在细胞核中以活化的脱磷酸化和失活的磷酸化的形式存在,活化的Rb蛋白对于细胞从G0/G1期进入S期有抑制作用,Rb基因失活,细胞就处于持续增殖期,并有可能恶变。细胞周期调控异常已经被证实为肿瘤发生发展过程中的一个重要机制,CyclinD1做为细胞周期G1-S期的重要调节因子之一,其表达的异常可使细胞进入增殖周期而发生异常增殖,最终导致肿瘤的发生。目的:该实验受试对象为人卵巢性乳头状囊腺癌SKOV3细胞,浆液给予IP6不同剂量单独应用及IP6中剂量与顺铂联合应用,观察SKOV3细胞的形态、数目等变化及Rb基因和CyclinD1蛋白的表达情况,探讨IP6对人卵巢浆液性乳头状囊腺癌SKOV3细胞的抑制作用,为卵巢癌的诊治和临床实践提供参考。方法:1细胞培养:在RPMI-1640培养基(含10%胎牛血清)中培养人卵巢癌SKOV3细胞系。将培养瓶放在37℃、含5%CO_2的培养箱中。2制备实验药物:实验药物均用0.9%氯化钠注射液配制成所需浓度,即配即用。3实验分组:培养基随机分为六组空白对照组,IP6低剂量组、IP6中剂量组、IP6高低剂量组,顺铂组,联合组(顺铂+IP6中剂量)。4实验所用药品浓度:空白对照组:新鲜培养基、IP6低剂量组1.5mmol/L、中剂量组3mmol/L、高剂量组6mmol/L、顺铂组3ug/ml,联合组浓度:顺铂:3ug/ml+IP6:3mmol/L。5 Real Time-PCR检测瘤组织中Rb基因的表达。6 Western Blot实验检测瘤组织中CyclinD1的表达情况。7统计学方法:采用SPSS13.0统计学软件对实验数据进行处理。以P0.05为差异具有统计学意义。结果:1卵巢癌细胞中Rb表达:所有分组均有Rb表达。在对照组中低表达,实验组中高表达,表达量的排列顺序为联合组顺铂组高剂量组中剂量组低剂量组对照组,差异具有统计学意义(P0.05)。2卵巢癌细胞中CyclinD1表达:所有分组中均有CyclinD1蛋白的表达,在对照组中高表达,实验组中低表达,表达量的排列顺序为联合组顺铂组高剂量组中剂量组低剂量组对照组,差异具有统计学意义(P0.05)。结论:1 IP6与顺铂均表现出对SKOV3细胞的抑制作用,且呈剂量依赖性;两者有协同作用。2 IP6增加Rb的表达,并降低CyclinD1表达,均表现出剂量依赖性。
[Abstract]:Ovarian cancer is one of the most common gynecological malignancy, its mortality rate ranks first in gynecological tumors, although many studies about ovarian cancer at home and abroad, but the cause of disease and its pathogenesis is not completely clear. Because the early symptoms of ovarian cancer is not easy to detect, advanced cases is also a lack of effective treatment, ascites and distant metastasis advanced symptoms have appeared most patients, although patients with advanced ovarian cancer can improve significantly the quality of life through surgery and chemotherapy, but in 2017 the U.S. National Cancer Institute survey: the five year survival rate was 45.6%[1] ovarian cancer, long-term survival rate is lower. Nowadays, ovarian cancer has become a serious threat to the life and health of women the main.2012 Nature gynecological cancer scientists found that some epithelial ovarian cancer originated from two transitional epithelial tissues, some originated from epithelial tissue stem [2] cells, therefore, many scholars are actively looking for the true origin of epithelial ovarian cancer. The study of ovarian cancer occurrence and development mechanism and explore the effective molecular marker and therapeutic target for ovarian cancer [3~4] is the research hotspot and difficulty of.IP6 (Inositol hexaphosphate) - six inositol phosphate, is a natural the phosphorus containing compound, its molecular structure includes inositol and phosphate ions. It takes 6 phytic acid (six inositol phosphate) and magnesium salt form rich in yeast, actinomyces, mammalian and plant [5], its anti-tumor effect has been confirmed in vivo and in vitro experiments, but the mechanism is not fully understood.IP6 has become a hot topic the research of anticancer drugs in recent years, and in colon cancer, lung cancer, breast cancer, but also to prove its efficient anticancer effect in vivo and in vitro experimental study on malignant tumor of prostate cancer and other types of IP6 at present. The main research contents include: immune to tumor cell killing effect, promote cancer cell apoptosis, anti mutation and DNA repair function and telomerase inhibition and antioxidant effect of [6-7].Rb gene is a tumor suppressor gene in activation and phosphorylation and inactivation of the phosphorylated form of activation in the nucleus. The Rb protein in cells from G0/G1 phase into S phase inhibition, inactivation of the Rb gene, cell proliferation on a sustained period, and may be malignant. Aberrant cell cycle regulation has been shown to occur as an important mechanism in the development of tumor, CyclinD1 as an important regulator of the G1-S phase of the cell cycle. The expression of the abnormal cells into the cell cycle and the occurrence of abnormal proliferation, eventually leading to the occurrence of cancer. Objective: the experimental subjects for human ovarian papillary serous cystadenocarcinoma SKOV3 cells to For the combined application of cisplatin dose and different doses of IP6 alone and IP6, observe the morphology of SKOV3 cells, the expression of the number of changes and Rb gene and CyclinD1 protein, to study the inhibitory effect of IP6 on human ovarian serous papillary cystadenocarcinoma of SKOV3 cells, and provide a reference for the diagnosis and treatment of ovarian cancer and clinical practice. Methods: 1 cell culture: in the RPMI-1640 medium (containing 10% fetal bovine serum) in cultured human ovarian cancer cell line SKOV3. The culture bottle at 37 degrees,.2 business incubator containing 5%CO_2 in the preparation of drugs: experimental drugs are used 0.9% Sodium Chloride Injection made the required concentration, which is equipped with that of.3 group: medium were randomly divided into six groups: control group, IP6 low dose group, middle dose of IP6 group, IP6 low dose group, cisplatin group, combination group (cisplatin +IP6) drug concentration by.4 experiment: control group: fresh culture medium, low dose group of IP6 1.5mmol/ L, 3mmol/L of middle dose group, high dose of 6mmol/L group, cisplatin group 3ug/ml, combined group: the concentration of cisplatin: expression of.7 CyclinD1 expression.6 Western statistical methods Blot assay of Rb gene detection of tumor tissue in Time-PCR 3ug/ml+IP6:3mmol/L.5 Real: the experimental data were processed by statistical software SPSS13.0. With P0.05 as statistically the significance of differences. Results: 1 ovarian cancer cells Rb expression: the expression of Rb. All groups were in control group and low expression, high expression in the experimental group, the expression of the order of the combined group cisplatin group high dose group, middle dose group and low dose group and control group, the difference was statistically significant (P0.05) expression of CyclinD1.2 in ovary cancer cells: showed the expression of CyclinD1 protein in all groups, the high expression in the control group, experimental group and low expression, the expression in the order of the joint group cisplatin group in high dose group The difference between the low dose group and the control group was statistically significant (P0.05). Conclusion: 1 IP6 and cisplatin both showed the inhibitory effect on SKOV3 cells in a dose-dependent manner. They had synergistic effects,.2 IP6 increased Rb expression and decreased CyclinD1 expression, all showed a dose-dependent manner.

【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.31

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