本文選題：二氫丹參酮I + 卵巢癌　； 參考：《大連醫(yī)科大學(xué)》2017年碩士論文

【摘要】：背景:卵巢癌是卵巢腫瘤的一種惡性腫瘤,其早期診斷比較困難,晚期不易治療,死亡率極高。目前,治療卵巢癌的有效手段是放療、化療和手術(shù)切除等。而化療是目前治療卵巢癌最廣泛,最主要的治療手段。臨床常用的化療藥物為順鉑,順鉑通過血液循環(huán)到達(dá)全身的組織和器官,從而造成DNA損傷殺死癌細(xì)胞。但順鉑對(duì)人體正常細(xì)胞也會(huì)產(chǎn)生一系列的細(xì)胞毒性,破壞人體的免疫系統(tǒng),此外,癌細(xì)胞會(huì)對(duì)順鉑產(chǎn)生耐藥性,因此,尋找一種高效且對(duì)正常細(xì)胞沒有毒性的抗卵巢癌藥物對(duì)卵巢癌的治療具有重要意義。二氫丹參酮I是丹參根莖的提取物,丹參是臨床中最常用的活血化瘀的中藥,相關(guān)文獻(xiàn)報(bào)道丹參具有增加冠脈血流量、擴(kuò)張冠狀動(dòng)脈、防止心肌缺血等心血管作用,以及用于治療胃癌癌、肝癌、宮頸癌等多種疾病。二氫丹參酮I是丹參中最主要的抗腫瘤成分,并且二氫丹參酮I對(duì)人體正常細(xì)胞毒性較小,但關(guān)于二氫丹參酮I抗腫瘤的作用機(jī)制尚不明確。目的:本研究主要是檢測(cè)二氫丹參酮I對(duì)卵巢癌的抑制作用機(jī)制,證明二氫丹參酮I抑制卵巢癌的增殖及轉(zhuǎn)移,為臨床治療卵巢癌提供了新的策略。方法:1.采用MTT法檢測(cè)體外培養(yǎng)的A2780和OV2008細(xì)胞的生長(zhǎng)與增殖情況。2.采用劃痕和侵襲實(shí)驗(yàn)檢測(cè)二氫丹參酮I對(duì)A2780和OV2008細(xì)胞轉(zhuǎn)移能力的影響。3.采用基因芯片技術(shù)檢測(cè)A2780細(xì)胞經(jīng)過二氫丹參酮I處理后的基因變化情況。4.根據(jù)目的基因PIK3CA的堿基序列,設(shè)計(jì)特異的過表達(dá)質(zhì)粒及無關(guān)(NC)序列,通過lipofectamin2000脂質(zhì)體法瞬時(shí)轉(zhuǎn)染A2780細(xì)胞,獲得PIK3CA表達(dá)上調(diào)的細(xì)胞模型,Western blot方法檢蛋白表達(dá)情況。5.q PCR和Western blot方法檢測(cè)細(xì)胞中目的基因的m RNA及蛋白質(zhì)水平的表達(dá)情況。6.采用MTT法和克隆形成實(shí)驗(yàn)比較二氫丹參酮I對(duì)A2780細(xì)胞與上調(diào)PIK3CA的A2780細(xì)胞生長(zhǎng)和增殖得變化情況。7.采用劃痕和侵襲實(shí)驗(yàn)比較二氫丹參酮I對(duì)A2780細(xì)胞與上調(diào)PIK3CA的A2780細(xì)胞的轉(zhuǎn)移能力。結(jié)果:1.MTT驗(yàn)證了二氫丹參酮I對(duì)A2780和OV2008細(xì)胞的增殖抑制作用,其抑制作用具有時(shí)間和劑量依賴關(guān)系。2.劃痕與侵襲實(shí)驗(yàn)證明二氫丹參酮I對(duì)A2780與OV2008的轉(zhuǎn)移具有抑制作用。3.基因芯片結(jié)果顯示經(jīng)二氫丹參酮I處理后A2780細(xì)胞中PIK3CA、DDB2、HGF等增殖與轉(zhuǎn)移相關(guān)基因下調(diào)。4.利用脂質(zhì)體瞬時(shí)轉(zhuǎn)染技術(shù),成功獲得PIK3CA表達(dá)上調(diào)的細(xì)胞模型,PIK3CA表達(dá)上調(diào)組細(xì)胞受二氫丹參酮I后的生長(zhǎng)和增殖抑制作用明顯低于對(duì)照組細(xì)胞。5.PIK3CA表達(dá)上調(diào)組細(xì)胞受二氫丹參酮I后的轉(zhuǎn)移抑制作用明顯低于對(duì)照組細(xì)胞。結(jié)論:研究結(jié)果表明二氫丹參酮I通過下調(diào)PIK3CA的表達(dá)影響PI3K/AKT信號(hào)通路抑制卵巢癌細(xì)胞的增殖與轉(zhuǎn)移。二氫丹參酮I治療癌癥具有潛在的臨床意義。
[Abstract]:Background: ovarian cancer is a malignant tumor of ovarian tumor, its early diagnosis is difficult, the late stage is difficult to treat, the mortality rate is extremely high. At present, the effective treatment of ovarian cancer is radiotherapy, chemotherapy and surgical resection. Chemotherapy is the most extensive and main treatment for ovarian cancer. The commonly used chemotherapeutic drug is cisplatin, which passes through the blood circulation to the tissues and organs of the body, causing DNA damage to kill cancer cells. But cisplatin also produces a series of cytotoxicity to human normal cells, destroying the body's immune system. In addition, cancer cells develop resistance to cisplatin, so, It is important to find an effective anti-ovarian cancer drug with no toxicity to normal cells for the treatment of ovarian cancer. Dihydrotanshinone I is an extract from the rhizome of Salvia miltiorrhiza. Salvia miltiorrhiza is the most commonly used Chinese medicine for promoting blood circulation and removing blood stasis. It is reported that Salvia miltiorrhiza has cardiovascular effects such as increasing coronary blood flow, expanding coronary artery, preventing myocardial ischemia, etc. And for the treatment of gastric cancer, liver cancer, cervical cancer and other diseases. Dihydrotanshinone I is the most important antitumor component in Danshen, and the toxicity of dihydrotanshinone I to human normal cells is relatively small, but the mechanism of dihydrotanshinone I anti-tumor is unclear. Objective: to investigate the mechanism of dihydrotanshinone I inhibiting ovarian cancer, and to prove that dihydrotanshinone I inhibits the proliferation and metastasis of ovarian cancer and provides a new strategy for clinical treatment of ovarian cancer. Method 1: 1. The growth and proliferation of cultured A2780 and OV2008 cells were detected by MTT assay. The effect of dihydrotanshinone I on metastasis ability of A2780 and OV2008 cells was detected by scratch and invasion assay. The gene changes of A2780 cells treated with dihydrotanshinone I were detected by gene chip technique. According to the base sequence of the target gene PIK3CA, a specific overexpression plasmid and an unrelated sequence were designed. A2780 cells were transiently transfected with lipofectamin2000 liposome. A cell model with up-regulated expression of PIK3CA was obtained. The expression of protein was detected by Western blot. 5.q PCR and Western blot were used to detect the expression of m RNA and protein level of the target gene. 6. The effects of dihydrotanshinone I on the growth and proliferation of A2780 cells and PIK3CA up-regulated A2780 cells were compared by MTT assay and clone formation assay. The metastatic ability of dihydrotanshinone I on A2780 cells and PIK3CA up-regulated A2780 cells was compared by scratch and invasion assay. Results 1. The inhibitory effects of dihydrotanshinone I on the proliferation of A2780 and OV2008 cells were confirmed by MTT. The inhibitory effects were in a time-and dose-dependent manner. Scratch and invasion tests showed that dihydrotanshinone I inhibited the metastasis of A2780 and OV2008. The microarray results showed that the proliferation and metastasis related genes such as PIK3CAG DDB2HGF were down-regulated in A2780 cells treated with dihydrotanshinone I. Using liposome transient transfection technology, The growth and proliferation inhibitory effect of PIK3CA up-regulated cells after dihydrotanshinone I was significantly lower than that of control cells. 5. PIK3CA expression upregulation group was inhibited by dihydrotanshinone I metastasis. The effect was significantly lower than that in the control group. Conclusion: dihydrotanshinone I inhibits the proliferation and metastasis of ovarian cancer cells by down-regulating the expression of PIK3CA. Dihydrotanshinone I has potential clinical significance in the treatment of cancer.
【學(xué)位授予單位】：大連醫(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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