本文選題：二氫丹參酮I + 卵巢癌　； 參考：《大連醫(yī)科大學》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)到達全身的組織和器官,從而造成DNA損傷殺死癌細胞。但順鉑對人體正常細胞也會產生一系列的細胞毒性,破壞人體的免疫系統(tǒng),此外,癌細胞會對順鉑產生耐藥性,因此,尋找一種高效且對正常細胞沒有毒性的抗卵巢癌藥物對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,相關文獻報道丹參具有增加冠脈血流量、擴張冠狀動脈、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對人體正常細胞毒性較小,但關于二氫丹參酮I抗腫瘤的作用機制尚不明確。目的:本研究主要是檢測二氫丹參酮I對卵巢癌的抑制作用機制,證明二氫丹參酮I抑制卵巢癌的增殖及轉移,為臨床治療卵巢癌提供了新的策略。方法:1.采用MTT法檢測體外培養(yǎng)的A2780和OV2008細胞的生長與增殖情況。2.采用劃痕和侵襲實驗檢測二氫丹參酮I對A2780和OV2008細胞轉移能力的影響。3.采用基因芯片技術檢測A2780細胞經過二氫丹參酮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,設計特異的過表達質粒及無關(NC)序列,通過lipofectamin2000脂質體法瞬時轉染A2780細胞,獲得PIK3CA表達上調的細胞模型,Western blot方法檢蛋白表達情況。5.q PCR和Western blot方法檢測細胞中目的基因的m RNA及蛋白質水平的表達情況。6.采用MTT法和克隆形成實驗比較二氫丹參酮I對A2780細胞與上調PIK3CA的A2780細胞生長和增殖得變化情況。7.采用劃痕和侵襲實驗比較二氫丹參酮I對A2780細胞與上調PIK3CA的A2780細胞的轉移能力。結果:1.MTT驗證了二氫丹參酮I對A2780和OV2008細胞的增殖抑制作用,其抑制作用具有時間和劑量依賴關系。2.劃痕與侵襲實驗證明二氫丹參酮I對A2780與OV2008的轉移具有抑制作用。3.基因芯片結果顯示經二氫丹參酮I處理后A2780細胞中PIK3CA、DDB2、HGF等增殖與轉移相關基因下調。4.利用脂質體瞬時轉染技術,成功獲得PIK3CA表達上調的細胞模型,PIK3CA表達上調組細胞受二氫丹參酮I后的生長和增殖抑制作用明顯低于對照組細胞。5.PIK3CA表達上調組細胞受二氫丹參酮I后的轉移抑制作用明顯低于對照組細胞。結論:研究結果表明二氫丹參酮I通過下調PIK3CA的表達影響PI3K/AKT信號通路抑制卵巢癌細胞的增殖與轉移。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.
【學位授予單位】：大連醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R737.31

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1 姜國強;二氫丹參酮Ⅰ抑制卵巢癌增殖與轉移作用的機制研究[D];大連醫(yī)科大學;2017年

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