本文關(guān)鍵詞：IP6對(duì)卵巢癌SKOV3細(xì)胞株增殖凋亡的影響　出處：《河北醫(yī)科大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 卵巢癌 SKOV3 IP6 Rb CyclinD1

【摘要】：卵巢癌是婦科常見的惡性腫瘤之一,其死亡率居?jì)D科腫瘤的首位,雖然國(guó)內(nèi)外有關(guān)卵巢癌的研究很多,但它的發(fā)病原因和發(fā)病機(jī)制尚未完全研究清楚。因卵巢癌早期癥狀不易察覺,晚期病例也缺乏有效的治療手段,多數(shù)患者確診時(shí)已出現(xiàn)腹水及遠(yuǎn)處轉(zhuǎn)移等晚期癥狀,雖然通過手術(shù)和化療能夠明顯的改善晚期卵巢癌患者的生活質(zhì)量,但2017年美國(guó)國(guó)家癌癥研究所調(diào)查:卵巢癌的五年生存率為45.6%[1],長(zhǎng)期生存率更低�，F(xiàn)如今,卵巢癌已成為嚴(yán)重威脅廣大女性生命健康的最主要的婦科腫瘤。2012年Nature的科學(xué)家發(fā)現(xiàn),一些卵巢上皮癌起源于兩種上皮組織的過渡帶,另一些起源于上皮組織的干細(xì)胞[2],因此,各國(guó)學(xué)者正在積極尋找上皮性卵巢癌真正的起源。研究卵巢上皮性癌發(fā)生和發(fā)展的相關(guān)機(jī)制及探求有效的分子標(biāo)記物和治療靶點(diǎn)[3~4]仍是目前卵巢癌研究的熱點(diǎn)和難點(diǎn)。IP6(Inositol hexaphosphate)--六磷酸肌醇,是一種天然的含磷化合物,其分子結(jié)構(gòu)包括肌醇和6個(gè)磷酸離子。它以植酸(肌醇六磷酸)鈣鎂鹽的形式豐富存在于酵母、放線菌、哺乳動(dòng)物和植物[5],其抗癌作用已在體內(nèi)外實(shí)驗(yàn)中得到證實(shí),但作用機(jī)制尚不完全清楚。IP6已成為近年來抗癌藥物研究的熱點(diǎn),并在結(jié)腸癌,肺癌,乳腺癌,前列腺癌等多種類型的惡性腫瘤體內(nèi)體外實(shí)驗(yàn)研究中也證明其有效的抗癌作用。目前對(duì)IP6主要的研究?jī)?nèi)容包括:對(duì)腫瘤細(xì)胞的免疫殺傷作用、促進(jìn)癌細(xì)胞凋亡作用、抗突變及DNA修復(fù)作用及端粒酶的抑制作用及抗氧化作用等[6-7]。Rb基因是一種腫瘤抑制基因,在細(xì)胞核中以活化的脫磷酸化和失活的磷酸化的形式存在,活化的Rb蛋白對(duì)于細(xì)胞從G0/G1期進(jìn)入S期有抑制作用,Rb基因失活,細(xì)胞就處于持續(xù)增殖期,并有可能惡變。細(xì)胞周期調(diào)控異常已經(jīng)被證實(shí)為腫瘤發(fā)生發(fā)展過程中的一個(gè)重要機(jī)制,CyclinD1做為細(xì)胞周期G1-S期的重要調(diào)節(jié)因子之一,其表達(dá)的異常可使細(xì)胞進(jìn)入增殖周期而發(fā)生異常增殖,最終導(dǎo)致腫瘤的發(fā)生。目的:該實(shí)驗(yàn)受試對(duì)象為人卵巢性乳頭狀囊腺癌SKOV3細(xì)胞,漿液給予IP6不同劑量單獨(dú)應(yīng)用及IP6中劑量與順鉑聯(lián)合應(yīng)用,觀察SKOV3細(xì)胞的形態(tài)、數(shù)目等變化及Rb基因和CyclinD1蛋白的表達(dá)情況,探討IP6對(duì)人卵巢漿液性乳頭狀囊腺癌SKOV3細(xì)胞的抑制作用,為卵巢癌的診治和臨床實(shí)踐提供參考。方法:1細(xì)胞培養(yǎng):在RPMI-1640培養(yǎng)基(含10%胎牛血清)中培養(yǎng)人卵巢癌SKOV3細(xì)胞系。將培養(yǎng)瓶放在37℃、含5%CO_2的培養(yǎng)箱中。2制備實(shí)驗(yàn)藥物:實(shí)驗(yàn)藥物均用0.9%氯化鈉注射液配制成所需濃度,即配即用。3實(shí)驗(yàn)分組:培養(yǎng)基隨機(jī)分為六組空白對(duì)照組,IP6低劑量組、IP6中劑量組、IP6高低劑量組,順鉑組,聯(lián)合組(順鉑+IP6中劑量)。4實(shí)驗(yàn)所用藥品濃度:空白對(duì)照組:新鮮培養(yǎng)基、IP6低劑量組1.5mmol/L、中劑量組3mmol/L、高劑量組6mmol/L、順鉑組3ug/ml,聯(lián)合組濃度:順鉑:3ug/ml+IP6:3mmol/L。5 Real Time-PCR檢測(cè)瘤組織中Rb基因的表達(dá)。6 Western Blot實(shí)驗(yàn)檢測(cè)瘤組織中CyclinD1的表達(dá)情況。7統(tǒng)計(jì)學(xué)方法:采用SPSS13.0統(tǒng)計(jì)學(xué)軟件對(duì)實(shí)驗(yàn)數(shù)據(jù)進(jìn)行處理。以P0.05為差異具有統(tǒng)計(jì)學(xué)意義。結(jié)果:1卵巢癌細(xì)胞中Rb表達(dá):所有分組均有Rb表達(dá)。在對(duì)照組中低表達(dá),實(shí)驗(yàn)組中高表達(dá),表達(dá)量的排列順序?yàn)槁?lián)合組順鉑組高劑量組中劑量組低劑量組對(duì)照組,差異具有統(tǒng)計(jì)學(xué)意義(P0.05)。2卵巢癌細(xì)胞中CyclinD1表達(dá):所有分組中均有CyclinD1蛋白的表達(dá),在對(duì)照組中高表達(dá),實(shí)驗(yàn)組中低表達(dá),表達(dá)量的排列順序?yàn)槁?lián)合組順鉑組高劑量組中劑量組低劑量組對(duì)照組,差異具有統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論:1 IP6與順鉑均表現(xiàn)出對(duì)SKOV3細(xì)胞的抑制作用,且呈劑量依賴性;兩者有協(xié)同作用。2 IP6增加Rb的表達(dá),并降低CyclinD1表達(dá),均表現(xiàn)出劑量依賴性。
[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.

【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R737.31

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