發(fā)布時(shí)間：2018-03-28 21:00

  本文選題：卵巢癌　切入點(diǎn)：SKOV3　出處：《河北醫(yī)科大學(xué)》2017年碩士論文

【摘要】：在婦科惡性腫瘤中,死亡率第一位的是卵巢癌,組織分型中以上皮性腫瘤占85%-90%,由于卵巢癌早期無特異性癥狀,且缺乏高敏感性及高特異性的篩查手段以及特效的治療方法,而傳統(tǒng)的化療方法存在許多副反應(yīng)以及化療藥物耐藥的問題,使卵巢癌的生存率仍然很低。天然藥物具有經(jīng)濟(jì)、高效、來源廣、低毒副作用等優(yōu)點(diǎn)。近年來,以肌醇六磷酸(IP6)為代表的植物提取的天然藥物以其強(qiáng)大的抗腫瘤作用成為近來研究的熱點(diǎn)。Wnt通路近來被廣泛關(guān)注,其可調(diào)控動(dòng)物組織分化、器官形成等生理過程,通路中的關(guān)鍵蛋白表達(dá)異常,可導(dǎo)致癌癥的發(fā)生。其中β-連環(huán)蛋白(β-catenin)與T細(xì)胞因子4(TCF4)都是Wnt通路中的主要成員。有研究表明卵巢良性疾病的β-catenin低于卵巢惡性疾病,并且β-catenin的異常表達(dá)與卵巢癌的臨床分期、病理分型、血行轉(zhuǎn)移情況有關(guān)。TCF4與β-catenin結(jié)合可以促進(jìn)基因轉(zhuǎn)錄,許多腫瘤的發(fā)生發(fā)展均與TCF4有關(guān)。上皮間質(zhì)轉(zhuǎn)化過程(epithelial mesenchymal transition,EMT)是上皮細(xì)胞向間質(zhì)細(xì)胞轉(zhuǎn)變的過程。EMT被認(rèn)為是腫瘤轉(zhuǎn)移的主要原因。Wnt經(jīng)典通路與EMT發(fā)生關(guān)聯(lián)密切,β-catenin、TCF4均與癌細(xì)胞EMT的發(fā)生以及腫瘤侵襲轉(zhuǎn)移密切相關(guān)。目的:本研究旨在探討IP6+肌醇對(duì)卵巢癌SKOV3細(xì)胞侵襲轉(zhuǎn)移能力的影響及其作用機(jī)制,以及順鉑與IP6+肌醇聯(lián)合應(yīng)用對(duì)卵巢癌是否有協(xié)同作用。方法:1 Transwell腫瘤細(xì)胞侵襲實(shí)驗(yàn),觀察IP6+肌醇劑量變化對(duì)卵巢癌細(xì)胞侵襲轉(zhuǎn)移能力的影響。2 Western blot法檢測(cè)IP6+肌醇對(duì)β-catenin蛋白的影響。3 RealtimePCR技術(shù)檢測(cè)Wnt通路中β-catenin mRNA,TCF4mRNA表達(dá)的影響。4統(tǒng)計(jì)學(xué)方法:采用SPSS21.0統(tǒng)計(jì)學(xué)軟件處理實(shí)驗(yàn)數(shù)據(jù)。結(jié)果:1transwell侵襲實(shí)驗(yàn)結(jié)果顯示:聯(lián)合組細(xì)胞數(shù)為26.2±2.6,順鉑組細(xì)胞數(shù)為32.0±3.4,聯(lián)合組、順鉑組與對(duì)照組82.8±5.7相比細(xì)胞數(shù)明顯減少,差異顯著(p0.05),ip6+肌醇高、中劑量組細(xì)胞數(shù)分別為62.6±2.3,73.2±3.0,與對(duì)照組相比,差異有統(tǒng)計(jì)學(xué)意義(p0.05)。ip6+肌醇低劑量組細(xì)胞數(shù)為80.2±3.3,與對(duì)照組相比,差異無統(tǒng)計(jì)學(xué)意義(p0.05),ip6+肌醇各濃度組中,隨著ip6濃度增高細(xì)胞數(shù)量減少,差異有統(tǒng)計(jì)學(xué)意義(p0.05)。2westernblot檢測(cè)β-catenin蛋白表達(dá):ip6+肌醇高、中劑量組分別為0.886±0.262,0.987±0.707,與對(duì)照組1.291±0.431相比,差異有統(tǒng)計(jì)學(xué)意義(p0.05)。ip6+肌醇低劑量組為1.245±0.580,與對(duì)照組相比無明顯差異(p0.05)。隨著ip6+肌醇劑量的增高,各組蛋白表達(dá)量逐漸降低,聯(lián)合組蛋白表達(dá)量為0.601±0.019,順鉑組為0.694±0.039,與對(duì)照組相比,差異有統(tǒng)計(jì)學(xué)意義(p0.05)。且聯(lián)合組蛋白表達(dá)量低于順鉑組,差異有統(tǒng)計(jì)學(xué)意義(p0.05)。3rt-pcr檢測(cè)β-cateninmrna的表達(dá):對(duì)照組為1.000±0.000,ip6+肌醇低劑量0.984±0.103,兩組無明顯差異(p0.05),ip6中劑量組、高劑量組、順鉑組、聯(lián)合組分別為0.906±0.010,0.786±0.104,0.570±0.123,0.488±0.126,與對(duì)照組相比,差異均有統(tǒng)計(jì)學(xué)意義(p0.05),聯(lián)合用藥組表達(dá)量低于順鉑組,差異有統(tǒng)計(jì)學(xué)意義(p0.05)。ip6各劑量組間,隨著劑量的增加,表達(dá)逐漸減弱,差異有統(tǒng)計(jì)學(xué)意義(p0.05)。4rt-pcr檢測(cè)tcf4mrna的表達(dá):ip6+肌醇高劑量組表達(dá)量為0.793±0.341,中劑量組為0.853±0.148,低劑量組為0.985±0.133,與對(duì)照組1.000±0.000相比,ip6各劑量組表達(dá)均降低,但與低劑量組差異不明顯(p0.05),與高、中劑量組相比,差異有統(tǒng)計(jì)學(xué)意義(p0.05)。隨著ip6劑量的升高,mrna的表達(dá)逐漸減弱。結(jié)論:1ip6+肌醇能抑制卵巢癌的侵襲轉(zhuǎn)移。2ip6+肌醇與順鉑具有協(xié)同抗癌作用。3ip6+肌醇阻礙卵巢癌侵襲轉(zhuǎn)移的機(jī)制可能是影響wnt信號(hào)通路,抑制腫瘤的emt,最終抑制其侵襲轉(zhuǎn)移。
[Abstract]:In gynecological malignant tumors, the first mortality is ovarian cancer, histological type of tumor of the skin above accounted for 85%-90%, due to early ovarian cancer specific symptoms, and the lack of means of screening Gao Min perceptual and high specificity and treatment effect, and the method of traditional chemotherapy has many side effects and chemotherapy the problem of drug resistance, the survival rate of ovarian cancer is still very low. The natural medicine is economic, efficient, wide source, low toxicity and other advantages. In recent years, with the inositol six phosphate (IP6) as the representative of the natural medicine plant extraction becomes a hot topic in recent research of.Wnt pathway has been extensively concerned with its anti-tumor effect strong, the regulation of animal tissue differentiation, organ formation and other physiological processes, the abnormal expression of key proteins in the pathway, there may lead to cancer. The beta catenin (beta -catenin) and T cell factor 4 (TCF4) is Key members of Wnt pathway. Studies have shown that ovarian benign disease beta -catenin was lower than that of ovarian malignant disease, and clinical abnormal expression in ovarian cancer and beta -catenin staging, pathological type, metastasis related.TCF4 and beta -catenin binding can promote gene transcription, occurrence and development of many tumors are associated with TCF4. Epithelial mesenchymal transition process (epithelial mesenchymal transition, EMT) is epithelial to mesenchymal transition.EMT is considered the main cause of tumor metastasis.Wnt classic pathway and EMT associated closely, beta -catenin, TCF4 and EMT cancer cells and tumor invasion and metastasis. Objective: This study aimed to investigate the effect of inositol IP6+ on invasion and metastasis of ovarian cancer SKOV3 cells and its mechanism, and the combination of cisplatin and IP6+ inositol on ovarian cancer have synergistic effect. Methods: 1 Tra Nswell tumor cell invasion assay, to observe the effect of.2 Western blot IP6+ inositol dose changes on invasion and metastasis of ovarian cancer cells to detect IP6+ inositol effect on beta -catenin protein.3 RealtimePCR detection of beta -catenin Wnt pathway mRNA, TCF4mRNA expression of.4 statistical methods: SPSS21.0 statistical software with experimental data. Results: 1transwell experimental results show that the combined group invasion cell number was 26.2 + 2.6, cisplatin group cells was 32 + 3.4, 82.8 + group, cisplatin group and control group compared to 5.7 cells significantly reduced the number of significant differences (P0.05), ip6+ high dose group of inositol, cell numbers were 62.6 + 2.3,73.2 + 3. Compared with the control group, the difference was statistically significant (P0.05).Ip6+ inositol low dose group of cells was 80.2 + 3.3, compared with the control group, the difference was not statistically significant (P0.05), ip6+ muscle alcohol each concentration group, with IP6 concentration The degree of increased number of cells decreased, the difference was statistically significant (P0.05) expression of beta -catenin protein.2westernblot: ip6+ inositol high, medium dose group were 0.886 + 0.262,0.987 + 0.707, 1.291 + 0.431 compared with the control group, the difference was statistically significant (P0.05).Ip6+ inositol low dose group was 1.245 + 0.580, compared with the control group no significant difference (P0.05). With the increase of ip6+ inositol dose, each protein expression decreased gradually, the combined group protein expression was 0.601 + 0.019, 0.694 + cisplatin group was 0.039, compared with the control group, the difference was statistically significant (P0.05). The expression of protein is lower than the combined group and cisplatin group, was statistically significant the difference (P0.05) to detect the expression of.3rt-pcr beta -cateninmrna: the control group is 1 + 0, 0.984 + 0.103 ip6+ inositol low dose, no significant difference between the two groups (P0.05), IP6 middle dose group, high dose group, cisplatin group and combination group were 0.906 + 0 .010,0.786 + 0.104,0.570 + 0.123,0.488 + 0.126, compared with the control group, the differences were statistically significant (P0.05), combination group expression was lower than that of cisplatin group, the difference was statistically significant (P0.05.Ip6) between the groups of different dose, with the dose increased, the expression decreased gradually, and there was statistically significant difference (P0.05) to detect the expression of.4rt-pcr tcf4mrna the high dose group of inositol: ip6+ expression level was 0.793 + 0.341, 0.853 + 0.148 in middle dose group, low dose group was 0.985 + 0.133, 1 + 0 compared with the control group, the expression of IP6 in each dose group were decreased, but with the low dose group was not significantly different (P0.05), and high dose. Compared with the group, the difference was statistically significant (P0.05). With the increase of IP6 dose, the expression of mRNA decreased. Conclusion: 1ip6+ inositol can inhibit ovarian cancer invasion and metastasis of.2ip6+ and cisplatin have synergistic anticancer effect of inositol inositol.3ip6+ hinder the invasion and metastasis of ovarian cancer machine It may affect the Wnt signaling pathway, inhibit the tumor's EMT, and eventually inhibit its invasion and metastasis.

【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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