本文選題：阿霉素 + HOE-140��； 參考：《河北醫(yī)科大學(xué)》2014年碩士論文

【摘要】：目的:卵巢惡性腫瘤是女性生殖器官常見的三大惡性腫瘤之一，由于卵巢位于盆腔深部，早期病變不易發(fā)現(xiàn)，晚期病例也缺乏有效的治療手段，因此卵巢惡性腫瘤致死率居?jì)D科惡性腫瘤首位，已成為嚴(yán)重威脅婦女生命和健康的主要腫瘤。卵巢癌的組織類型繁多，其中上皮性卵巢癌占卵巢惡性腫瘤的85~90%，其發(fā)病隱匿，早期無癥狀，發(fā)現(xiàn)時(shí)往往已是晚期，這使得該腫瘤的早診早治顯得異常困難。 對(duì)于卵巢癌的治療，適當(dāng)?shù)氖中g(shù)配合有效化療可大大提高病人的存活率，其中上皮性卵巢癌對(duì)化療藥物較敏感。其傳統(tǒng)的化療方法是紫杉醇加鉑類，但是多數(shù)患者停藥一段時(shí)間后會(huì)復(fù)發(fā)，這可能和卵巢癌的發(fā)病機(jī)制尚不清楚和腫瘤耐藥有關(guān)。 緩激肽(BK)是機(jī)體炎癥時(shí)釋放的一種血管活性肽，，通過和細(xì)胞膜上的B l受體和B2受體結(jié)合而起效。對(duì)卵巢癌、胃癌和肝癌等多種病人的胸腹腔滲液檢查均發(fā)現(xiàn)高濃度的BK水平，對(duì)其受體的研究發(fā)現(xiàn)，B1受體和B2受體單獨(dú)或同時(shí)在人類多種腫瘤細(xì)胞呈高表達(dá)。這些研究提示，B1R、B2R或二者均可參與腫瘤的增殖。但BK受體亞型在卵巢癌組織的表達(dá)尚未有一致報(bào)道。 在本部分研究中，我們首先就上皮性卵巢癌患者和卵巢良性腫瘤患者經(jīng)手術(shù)切除的腫塊組織中BK受體亞型及其下游的主要信號(hào)分子NO產(chǎn)生的酶eNOS和iNOS的表達(dá)進(jìn)行了比較研究。在此基礎(chǔ)上，觀察了B2受體的特異性拮抗劑HOE-140單獨(dú)或聯(lián)合應(yīng)用阿霉素（DOX）對(duì)人卵巢癌細(xì)胞SKOV3的影響及可能的作用機(jī)制。該研究有望找到更好的腫瘤靶位，從而改善卵巢癌患者的生存質(zhì)量。 方法： 1卵巢癌組織中緩激肽B1受體、B2受體及eEOS、iNOS的表達(dá) 選取臨床確診的卵巢癌患者及卵巢上皮性良性腫瘤患者各20例。年齡在35~55歲之間。無高血壓、糖尿病等內(nèi)科疾病及遺傳病病史。 采用患者手術(shù)切除后經(jīng)病理確診為卵巢上皮性良性腫瘤及上皮性卵巢癌患者的卵巢標(biāo)本，免疫組化法檢測卵巢癌組織中B1、B2及eNOS及iNOS的表達(dá)。 2血清中NO活性測定 在治療前抽取初步診斷為卵巢癌的患者靜脈血液5ml20例，（其中15例術(shù)后病理證實(shí)為上皮性卵巢癌），另抽取正常人靜脈血液5ml20例，離心，分離上清液，采用NO試劑盒測定血液中NO含量。 3DOX和HOE-140單獨(dú)或聯(lián)合應(yīng)用對(duì)SKOV3細(xì)胞增值的影響 采用MTT法檢測不同濃度的阿霉素（3,10,30,100和300μmol/L）或HOE-140(0.01,0.1,1,10和100μmol/L)和SKOV3作用24小時(shí)后細(xì)胞的存活情況。在此基礎(chǔ)上，比較二者聯(lián)合用藥（DOX10μmol/L+HOE14010μmol/L）和分別單獨(dú)應(yīng)用相同劑量的DOX或HOE-140對(duì)SKOV3的細(xì)胞毒作用。 4DOX和HOE-140單獨(dú)或聯(lián)合應(yīng)用對(duì)SKOV3細(xì)胞凋亡的影響 將SKOV3細(xì)胞分別與DOX10μmol/L、HOE-14010μmol/L或聯(lián)合應(yīng)用DOX10μmol/L+HOE-14010μmol/L作用24h，采用原位末端標(biāo)記技術(shù)觀察(TUNEL試劑盒)細(xì)胞凋亡情況。 結(jié)果： 1卵巢組織標(biāo)本B1、B2受體、eEOS和iNOS的表達(dá) 對(duì)臨床上確診為卵巢癌患者和卵巢良性腫瘤患者手術(shù)后切取的卵巢組織進(jìn)行免疫組化結(jié)果顯示，B1受體在良性卵巢組織中幾乎不表達(dá)，B2受體在良性卵巢組織中有表達(dá)，eNOS在良性卵巢組織中有少量表達(dá)，iNOS無明顯表達(dá)，和卵巢良性腫瘤患者的卵巢組織相比，卵巢癌組織細(xì)胞中B1受體、B2受體、eNOS及iNOS均呈高表達(dá)。 2血清中NO含量測定 對(duì)所采集血清中NO含量測定結(jié)果表明，卵巢癌患者血清中NO含量顯著高于正常人血清中的含量（P0.01）。 3對(duì)SKOV3細(xì)胞增殖的影響 MTT法檢測結(jié)果顯示，將不同濃度的DOX（3,10,30,100和300μmol/L）或HOE-140(0.01,0.1,1,10和100μmol/L)與SKOV3共同孵育24小時(shí)后，二者均對(duì)SKOV3細(xì)胞的增殖有一定的抑制作用。其中HOE-14010μmol/L的抑制率是12.7%，而DOX10μmol/L的抑制率為26.8%。二者聯(lián)合應(yīng)用的抑制率為37.5%，其聯(lián)合應(yīng)用的對(duì)SOV3細(xì)胞的抑制作用明顯強(qiáng)于二者單獨(dú)應(yīng)用（P均0.01）。 4對(duì)SKOV3細(xì)胞凋亡的影響 Tunnel染色顯示，單獨(dú)應(yīng)用10μmol/L HOE-140或DOX處理SKOV3細(xì)胞24小時(shí)均可誘導(dǎo)SKOV3細(xì)胞的凋亡，兩者聯(lián)合應(yīng)用誘導(dǎo)SKOV3細(xì)胞的凋亡作用明顯強(qiáng)于單獨(dú)應(yīng)用。 結(jié)論： 1上皮性卵巢癌患者血清中NO含量較正常人為高，其卵巢組織中B1、B2、eNOS及iNOS的表達(dá)都增加; 2HOE-140和DOX聯(lián)合或單獨(dú)應(yīng)用均可抑制卵巢癌SKOV3細(xì)胞的增殖，其機(jī)制可能和促進(jìn)細(xì)胞凋亡和抑制NO產(chǎn)生有關(guān)。
[Abstract]:Objective: ovarian malignant tumor is one of the three common malignant tumors in female genital organs. Because the ovary is located in the deep pelvic cavity, the early pathological changes are not easy to be found, and the late cases are also lack of effective treatment. Therefore, the mortality rate of ovarian malignant tumor is the first one in gynecologic malignant tumor, and has become the major tumor that seriously threatens the life and health of women. There are various types of tissue in ovarian cancer, among which epithelial ovarian cancer accounts for 85~90% of ovarian malignant tumors. The pathogenesis is occult, early asymptomatic, and often late in discovery, which makes the early diagnosis of the tumor very difficult.
For the treatment of ovarian cancer, proper operation and effective chemotherapy can greatly improve the patient's survival rate, of which epithelial ovarian cancer is more sensitive to chemotherapeutic drugs. The traditional chemotherapy method is paclitaxel plus platinum, but most patients relapse after a period of withdrawal, which may not be known to the pathogenesis of ovarian cancer and the tumor resistance. Of
Bradykinin (BK) is a vasoactive peptide released during the inflammation of the body, which is effective by combining with the B l receptor and B2 receptor on the cell membrane. A high concentration of BK levels is found in a variety of patients with ovarian cancer, gastric cancer, and liver cancer, and the study of its receptors, B1 receptor and B2 receptor alone or at the same time in human multiple swollen. The tumor cells are highly expressed. These studies suggest that B1R, B2R, or two may participate in the proliferation of tumors. However, the expression of BK receptor subtypes in ovarian cancer has not yet been reported.
In this part, we compare the expression of the BK receptor subtype and the expression of the enzyme eNOS and iNOS produced by the main signal molecule NO downstream of the excised tumor and the patients with epithelial ovarian cancer and benign ovarian tumors. On this basis, the specific antagonist HOE-140 of the B2 receptor, HOE-140 alone or in conjunction, is observed. The effect of adriamycin (DOX) on human ovarian cancer cell SKOV3 and its possible mechanism of action. This study is expected to find a better tumor target and improve the quality of survival of ovarian cancer patients.
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本文編號(hào)：1963563