本文關(guān)鍵詞：精子相關(guān)抗原9（SPAG9）在宮頸癌及宮頸上皮內(nèi)瘤變中的表達(dá)及意義　出處：《山東大學(xué)》2014年碩士論文　論文類型：學(xué)位論文

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【摘要】：背景： 宮頸癌是女性第三常見(jiàn)的惡性腫瘤,在惡性腫瘤死亡率中占第四位,嚴(yán)重威脅女性健康。并且,全球超過(guò)85%宮頸癌新發(fā)及死亡病例發(fā)生于發(fā)展中國(guó)家。宮頸癌的發(fā)生與發(fā)展是一個(gè)多階段、多步驟、復(fù)雜的過(guò)程,從癌前病變發(fā)展為宮頸癌可經(jīng)歷十余年時(shí)間,故宮頸癌是一種可預(yù)防、甚至可治愈的疾病。盡管,近些年來(lái)國(guó)內(nèi)外逐漸普及宮頸癌的篩查工作,雖已使宮頸癌的發(fā)病率、死亡率有明顯下降,但是因?qū)m頸液基細(xì)胞學(xué)檢查特異性以及敏感性不高,并且陰道鏡和宮頸活檢等屬創(chuàng)傷性檢查,也存在漏診的可能。另外,宮頸病變的術(shù)前及預(yù)后監(jiān)測(cè)并不完善,目前尚沒(méi)有一種可靠的腫瘤標(biāo)志物檢測(cè)。因此,尋找一種能為宮頸病變的早期篩查、臨床診斷及術(shù)后檢測(cè)提供幫助的新型腫瘤標(biāo)志物顯得尤為重要。目的： 檢測(cè)精子相關(guān)抗原9(SPAG9)在宮頸癌、宮頸上皮內(nèi)瘤變(cervical intraepithelial neoplasia, CIN)組織中的表達(dá),并檢測(cè)SPAG9在宮頸病變患者血清內(nèi)的免疫原性,評(píng)估其作為新宮頸癌血清標(biāo)志物的潛在價(jià)值。方法： 1、研究對(duì)象：子宮頸病變組織及正常宮頸組織選取自山東大學(xué)附屬省立醫(yī)院2009年至2012年宮頸手術(shù)或活檢患者標(biāo)本。新鮮子宮頸癌標(biāo)本40例；宮頸上皮內(nèi)瘤變(CIN)患者手術(shù)后石蠟標(biāo)本35例、新鮮標(biāo)本7例；對(duì)照組選取20例正常宮頸組織(因非宮頸病變而手術(shù))。另外,40例子宮頸癌、40例CIN患者血樣均取自須行手術(shù)患者,另收集20例子宮頸癌術(shù)后返院化療患者、20例子宮肌瘤患者及10例健康女性血樣做為對(duì)照。 2、研究方法： (1)應(yīng)用免疫組織化學(xué)(IFC)方法,明確SPAG9在各宮頸組織中的表達(dá)定位,通過(guò)對(duì)平均光密度值的檢測(cè),比較正常宮頸組織、宮頸上皮內(nèi)瘤變組織以及子宮頸癌組織中SPAG9的表達(dá)差異。 (2)應(yīng)用逆轉(zhuǎn)錄-聚合酶鏈反應(yīng)(RT-PCR)方法檢測(cè)正常宮頸組織、CIN組織及宮頸癌組織中SPAG9的表達(dá),并通過(guò)半定量分析各組間差異。 (3)通過(guò)Western Blot方法檢測(cè)正常宮頸組織、CIN組織及宮頸癌組織中SPAG9的蛋白表達(dá),進(jìn)行半定量分析。 (4)應(yīng)用酶聯(lián)免疫吸附試驗(yàn)(ELISA)檢測(cè)10例健康女性、20例子宮肌瘤患者、40例宮頸上皮內(nèi)瘤變患者、40例宮頸癌患者以及20例宮頸癌術(shù)后返院化療、無(wú)復(fù)發(fā)患者的血清標(biāo)本中SPAG9濃度差異。 結(jié)果： 1、免疫組織化學(xué)結(jié)果顯示,SPAG9主要定位于腫瘤細(xì)胞的細(xì)胞質(zhì)和細(xì)胞膜上,間質(zhì)中則極少量表達(dá)或不表達(dá)。SPAG9在正常宮頸組、宮頸上皮內(nèi)瘤變組及宮頸癌組中的表達(dá)量依次增強(qiáng),且各組間表達(dá)的差異均有統(tǒng)計(jì)學(xué)意義。但SPAG9的表達(dá)與宮頸癌臨床病理學(xué)特征,無(wú)論在病理類型、組織學(xué)分型、臨床分期或有無(wú)淋巴結(jié)轉(zhuǎn)移等方面的表達(dá)差異均無(wú)統(tǒng)計(jì)學(xué)意義。 2、RT-PCR結(jié)果顯示SPAG9在正常宮頸組織中極少表達(dá)或無(wú)表達(dá),而在宮頸癌組織中表達(dá)量明顯增高。通過(guò)對(duì)SPAG9mRNA在不同宮頸組織中表達(dá)的半定量分析顯示,在宮頸癌組織中平均表達(dá)量為0.815±0.553,而在正常宮頸組織中為0.008±0.001,兩組之間差異具有統(tǒng)計(jì)學(xué)意義。 3、Western Blot檢測(cè)SPAG9蛋白在宮頸癌及正常宮頸組織中的表達(dá)趨勢(shì),與免疫組織化學(xué)以及RT-PCR的結(jié)果一致。半定量分析SPAG9蛋白在宮頸癌組織中平均表達(dá)量為0.929±0.908,正常宮頸組織中平均表達(dá)量?jī)H為0.044±0.032,差異有統(tǒng)計(jì)學(xué)意義。 4、ELISA結(jié)果示10名健康女性血清中SPAG9平均血清濃度3.2343±1.0579ng/mL,20例子宮肌瘤患者中為3.3561±0.7426ng/mL,兩組間差異無(wú)統(tǒng)計(jì)學(xué)意義。而在40例宮頸上皮內(nèi)瘤變及40例宮頸癌患者血清中SPAG9的平均表達(dá)量分別為6.7015±3.3101ng/mL、15.1404±10.3839ng/mL,宮頸癌組高于CIN組高于子宮肌瘤及健康女性組,且各組間的差異均有統(tǒng)計(jì)學(xué)意義。另外,在20例宮頸癌術(shù)后返院化療、且無(wú)復(fù)發(fā)的患者血清中SPAG9平均濃度為3.6107±1.5069,較術(shù)前有所下降。結(jié)論： 1、在宮頸癌和CIN組織中SPAG9的蛋白和mRNA均有高表達(dá),且宮頸癌組高于CIN組高于正常宮頸組,三組間差異均有統(tǒng)計(jì)學(xué)意義。 2、宮頸癌及CIN患者血清中SPAG9表達(dá)量均高于正常健康女性和子宮肌瘤患者,且宮頸癌術(shù)后、未復(fù)發(fā)患者血清濃度較術(shù)前有所下降。 3、SPAG9作為一個(gè)新的生物學(xué)標(biāo)志物,可為宮頸癌及宮頸上皮內(nèi)瘤變的早期診斷、術(shù)后檢測(cè)、隨診治療提供指導(dǎo)意義。
[Abstract]:Background:
Cervical cancer is the third most common female malignant tumors, accounting for fourth in the mortality of malignant tumor, a serious threat to women's health. And, more than 85% of cervical cancer and deaths occur in developing countries. The occurrence and development of cervical cancer is a multi-stage, multi-step and complex process, from the development of precancerous lesions for cervical cancer can be more than ten years of experience in the Imperial Palace, cervical cancer is a preventable and treatable disease. Even though, in recent years at home and abroad gradually spread screening of cervical cancer, although the incidence of cervical cancer, mortality rate decreased significantly, but due to cervical cytology specificity and sensitivity is not high, and colposcopy and cervical biopsy is a traumatic examination, there is the possibility of missed diagnosis. In addition, cervical lesions before surgery and prognosis monitoring is not perfect, there is not a reliable tumor marker detection. Therefore, it is very important to find a new type of tumor marker for early screening of cervical lesions, clinical diagnosis and postoperative detection.
Detection of sperm associated antigen 9 (SPAG9) in cervical cancer, cervical intraepithelial neoplasia (cervical intraepithelial, neoplasia, CIN) expression, and detection of SPAG9 in serum of patients with cervical lesions in immunogenicity, evaluate its potential value as a new cervical cancer serum markers:
1, the object of study: cervical lesions and normal cervical tissue were selected from the affiliated Provincial Hospital of Shandong University from 2009 to 2012. Cervical biopsy or surgical specimens from patients with fresh cervical cancer specimens in 40 cases; cervical intraepithelial neoplasia (CIN) in 35 cases of paraffin embedded specimens of patients after surgery, 7 cases of fresh specimens from 20 cases of normal control group; the cervical tissue (for non cervical lesions and surgery). In addition, 40 cases of cervical carcinoma, 40 cases of CIN patients blood samples were taken from the patients with surgery should be performed, another collection of chemotherapy of 20 cervical cancer patients after hospital, 20 cases of uterine leiomyoma and 10 cases of healthy women were as control.
2, research methods:
(1) immunohistochemical localization (SPAG9) was applied to identify the expression and location of IFC in cervical tissues. The expression of SPAG9 in normal cervical tissues, cervical intraepithelial neoplasia tissues and cervical cancer tissues was compared by measuring the mean optical density.
(2) reverse transcription polymerase chain reaction (RT-PCR) was used to detect the expression of SPAG9 in normal cervical tissues, CIN tissues and cervical cancer tissues.
(3) the Western Blot method was used to detect the protein expression of SPAG9 in the normal cervical tissues, CIN tissues and cervical cancer tissues, and the semi quantitative analysis was performed.
(4) by enzyme linked immunosorbent assay (ELISA) was detected in 10 cases of healthy women, 20 patients with uterine myoma, 40 cases with cervical intraepithelial neoplasia, 40 cases of cervical cancer patients and 20 cases of cervical cancer hospital after chemotherapy, the SPAG9 concentration difference of serum samples in patients with no recurrence.
Result錛，

本文編號(hào)：1420987