本文選題：FR-α蛋白 + Ki-67��； 參考：《河北醫(yī)科大學(xué)》2014年碩士論文

【摘要】：目的：宮頸癌(Cervical Cancer)是最常見的婦科惡性腫瘤之一，發(fā)病率居女性惡性腫瘤的第二位,僅次于乳腺癌,其病理類型常見的有鱗癌、腺癌、腺鱗癌，其發(fā)病原因主要與以下因素有關(guān)：1.高危型HPV感染；2.性行為及分娩次數(shù)；3.沙眼衣原體、單純皰疹病毒II型、滴蟲等病原體的感染起著協(xié)同作用；4.吸煙以及營養(yǎng)不良、衛(wèi)生條件差等。發(fā)病早期的宮頸癌常無明顯的癥狀和體征，宮頸光滑或難與宮頸柱狀上皮異位相鑒別。頸管型患者因?qū)m頸外觀正常易漏診或誤診。其轉(zhuǎn)移途徑主要為直接蔓延及淋巴轉(zhuǎn)移，血行轉(zhuǎn)移較少見。在細(xì)胞代謝、DNA合成和修復(fù)以及癌細(xì)胞快速增殖分裂過程中葉酸發(fā)揮著重要作用，快速分裂癌細(xì)胞的DNA合成對于葉酸有很高的需求，所以近年來轉(zhuǎn)運(yùn)葉酸進(jìn)入細(xì)胞內(nèi)的葉酸受體α成為人們的研究熱點(diǎn)。葉酸受體是一種膜結(jié)合蛋白，它在結(jié)合運(yùn)輸生理水平的葉酸到細(xì)胞內(nèi)的過程中發(fā)揮著重要的作用，其在正常組織中低表達(dá)，在腫瘤細(xì)胞膜表面高度表達(dá)，葉酸受體（FR）分為FR-α、FR-β、FR-γ，其中FR-α高表達(dá)于黏液腺癌中，包括卵巢腺癌、子宮腺癌、宮頸腺癌、睪丸絨毛膜癌和某些腦腫瘤中，并且少量地表達(dá)于腎、結(jié)腸及心臟等癌癥中。Ki-67是存在于增殖細(xì)胞核的與細(xì)胞增殖密切相關(guān)的核蛋白，作為評價(jià)細(xì)胞增殖的標(biāo)記物，現(xiàn)已廣泛地應(yīng)用于腫瘤的研究和探討中。本實(shí)驗(yàn)用免疫組織化學(xué)法（immunohistochemistry）檢測宮頸癌組織中FR-α蛋白及Ki-67的表達(dá)，探討二者的關(guān)系及其臨床意義。 方法：采用免疫組織化學(xué)PV法檢測正常宮頸組織、宮頸上皮內(nèi)瘤變組織以及宮頸癌組織中FR-α蛋白表達(dá)及Ki-67的表達(dá)。分實(shí)驗(yàn)組：宮頸癌33例（其中宮頸癌I期11例，宮頸癌II期12例，，宮頸癌III期10例）。對照組：正常宮頸組織30例，宮頸上皮內(nèi)瘤變組織42例，陽性表達(dá)判斷：細(xì)胞胞漿內(nèi)出現(xiàn)棕黃色顆粒為準(zhǔn)，參考Soslow RA和劉祖國等[1][2]提到的方法，采用免疫組化評分法來判定結(jié)果，先按染色強(qiáng)度評分：未見染色為0分，淡黃色為1分,棕黃色為2分，棕褐色為3分；再按陽性細(xì)胞所占百分比評分：隨機(jī)選取高倍鏡（×200）下觀察5個(gè)視野，每個(gè)視野計(jì)數(shù)100個(gè)細(xì)胞，進(jìn)行陽性細(xì)胞數(shù)分級，0-1%=0，1-10%=1，10-50%=2，50-80%=3，80-100%=4)。兩項(xiàng)評分乘積所得總分（IHS）進(jìn)行結(jié)果判定：IHS0分為（-），IHS1~4分為（+），IHS5~8分為（++），IHS9~12分為（+++）。Ki-67結(jié)果判斷：細(xì)胞胞漿內(nèi)出現(xiàn)棕黃色顆粒為準(zhǔn)，參考Soslow RA和劉祖國等[1][2]提到的方法，采用免疫組化評分法來判定結(jié)果，先按染色強(qiáng)度評分：未見染色為0分，淡黃色為1分，棕黃色為2分，棕褐色為3分；再按陽性細(xì)胞所占百分比評分：隨機(jī)選取高倍鏡（×200）下觀察5個(gè)視野，每個(gè)視野計(jì)數(shù)100個(gè)細(xì)胞，行陽性細(xì)胞數(shù)分級，0-1%=0，1-10%=1，10-50%=2，50-80%=3，80-100%=4）。兩項(xiàng)評分乘積所得總分（IHS）進(jìn)行結(jié)果判定：IHS0分為（-），IHS1~4分為（+），IHS5~8分為（++），IHS9~12分為（+++）。所有數(shù)據(jù)采用SPSS13.0統(tǒng)計(jì)軟件處理。計(jì)數(shù)資料采用χ2（PearsonChi-Square Test）或Fish確切概率法；兩組變量間的相關(guān)分析采用Spearman等級相關(guān)分析。所有檢驗(yàn)均為雙側(cè)，P＜0.05為有統(tǒng)計(jì)學(xué)意義。 結(jié)果： 1正常宮頸組織、宮頸上皮內(nèi)瘤變組織、宮頸癌組織中FR-α蛋白表達(dá) 正常宮頸組織、宮頸上皮內(nèi)瘤變組織、宮頸癌組織中FR-α蛋白表達(dá)于細(xì)胞漿中，呈棕黃色顆粒。在本實(shí)驗(yàn)中，正常宮頸組織FR-α蛋白表達(dá)陽性率為6.7%，宮頸上皮內(nèi)瘤變組織FR-α蛋白表達(dá)陽性率為76.2%，宮頸癌組織FR-α蛋白表達(dá)陽性率為81.7%，差異有統(tǒng)計(jì)學(xué)意義（P＜0.05），見table1。宮頸上皮內(nèi)瘤變組織與正常組織中FR-α的表達(dá)陽性率相比，差異有統(tǒng)計(jì)學(xué)意義（P＜0.05），見table1。宮頸癌組織與宮頸上皮內(nèi)瘤變組織中FR-α蛋白表達(dá)陽性率相比，差異無統(tǒng)計(jì)學(xué)意義（P0.05），見table1。 2不同病理類型中FR-α蛋白表達(dá) CINI、CINII、CINIII中FR-α蛋白表達(dá)有差異，三者相比無統(tǒng)計(jì)學(xué)意義（P0.05）。見table2。宮頸癌I期、宮頸癌II期、宮頸癌III期蛋白表達(dá)有差異，三者差異無統(tǒng)計(jì)學(xué)意義（P0.05）。見Table3 3FR-α蛋白表達(dá)與宮頸癌危險(xiǎn)因素的相關(guān)性 在宮頸癌組織及宮頸上皮內(nèi)瘤變組織中，F(xiàn)R-α蛋白表達(dá)與年齡因素?zé)o關(guān),年齡40與年齡≤40兩組差別無統(tǒng)計(jì)學(xué)意義（P0.05），見Table4。在兩組中，F(xiàn)R-α蛋白表達(dá)與HPV感染均有關(guān),HPV感染陽性與HPV感染陰性兩組差別有統(tǒng)計(jì)學(xué)意義（P0.05），見Table4。 4正常宮頸組織、宮頸上皮內(nèi)瘤變組織、宮頸癌組織中Ki-67表達(dá) 正常宮頸組織、宮頸上皮內(nèi)瘤變組織、宮頸癌組織中Ki-67表達(dá)于細(xì)胞核中，呈棕黃色顆粒。在正常宮頸組織中未見Ki-67表達(dá)（見Fig.4），宮頸上皮內(nèi)瘤變組織及宮頸癌組織中Ki-67表達(dá)升高（見Fig.5,Fig.6），正常宮頸組織中Ki-67陽性率0%，宮頸上皮內(nèi)瘤變組織中Ki-67陽性率71.4%，宮頸癌病變組織中Ki-67陽性率84.8%，三者差異有統(tǒng)計(jì)學(xué)意義(P＜0.05)。見table5，正常宮頸組織與宮頸上皮內(nèi)瘤變組織中Ki-67蛋白表達(dá)陽性率相比，兩者差異有統(tǒng)計(jì)學(xué)意義（P0.05）,宮頸上皮內(nèi)瘤變組織與宮頸癌病變組織中Ki-67蛋白表達(dá)陽性率相比，兩者差異無統(tǒng)計(jì)學(xué)意義（P0.05）。見Table5。 5不同病理類型中Ki-67表達(dá) CINI、CINII、CINIII蛋白表達(dá)有差異，三者相比差異無統(tǒng)計(jì)學(xué)意義（P0.05）。見table6。宮頸癌I期、宮頸癌II期、宮頸癌III期Ki-67蛋白表達(dá)有差異，三者差異無統(tǒng)計(jì)學(xué)意義（P0.05），見Table7。 6Ki-67蛋白表達(dá)與宮頸癌危險(xiǎn)因素的相關(guān)性 在宮頸癌及宮頸上皮內(nèi)瘤變組織中，Ki-67蛋白表達(dá)與年齡因素?zé)o關(guān)，年齡40與年齡≤40兩組差別無統(tǒng)計(jì)學(xué)意義（P0.05），見Table8。在兩組中Ki-67蛋白表達(dá)與HPV感染無關(guān)，HPV感染陽性與HPV感染陰性兩組差別無統(tǒng)計(jì)學(xué)意義（P0.05）,見Table8。 7宮頸上皮內(nèi)瘤變組織及宮頸癌組織中FR-α蛋白表達(dá)與Ki-67的關(guān)系： 宮頸癌組織中二者呈正相關(guān)，相關(guān)系數(shù)r=0.532，p=0.000，有相關(guān)性，有統(tǒng)計(jì)學(xué)意義（P0.05)。宮頸上皮內(nèi)瘤變組織中，F(xiàn)R-α蛋白表達(dá)與Ki-67蛋白表達(dá)呈正相關(guān)，相關(guān)系數(shù)r=0.504，p=0.003，有相關(guān)性，有統(tǒng)計(jì)學(xué)意義（P0.05） 結(jié)論： 1FR-α蛋白表達(dá)升高參與了宮頸癌發(fā)病。 2FR-α蛋白表達(dá)與年齡因素?zé)o關(guān)，與HPV感染有協(xié)同作用。 3Ki-67的高表達(dá)促進(jìn)了腫瘤的發(fā)生，與HPV感染無協(xié)同作用。 4宮頸癌及宮頸上皮內(nèi)瘤變組織中FR-α蛋白表達(dá)與Ki-67有相關(guān)性；FR-α蛋白可能通過影響細(xì)胞增殖參與宮頸癌的發(fā)生與發(fā)展。
[Abstract]:Objective : Cervical cancer ( cervical cancer ) is one of the most common gynecological malignant tumors , the second place in the malignant tumor of breast cancer , which is second only to breast cancer . Its pathological type is squamous cell carcinoma , adenocarcinoma , adenosquamous cell carcinoma . The reason of the disease is related to the following factors : 1 . High - risk HPV infection ;
2 . Sex and number of deliveries ;
3 . The infection plays a synergistic role in the infection of the pathogen such as chlamydia , herpes simplex virus type II , trichomonad , etc .
The folate receptor is a kind of membrane - binding protein which plays an important role in the process of cell metabolism , DNA synthesis and repair and the rapid proliferation and division of cancer cells . The folate receptor is a kind of membrane - binding protein .

Methods : The expression of FR - 偽 protein and the expression of Ki - 67 in normal cervical tissue , cervical epithelial neoplasia and cervical cancer were detected by immunohistochemical method . In the control group , 30 cases of normal cervical tissue , 42 cases of cervical intraepithelioma and 42 cases of positive expression were determined : brown - yellow granules appeared in the cytoplasm of the cells , and the results were determined by immunohistochemical method .
The percentage of positive cells was scored as percentage of positive cells : 5 visual fields were randomly selected , 100 cells were counted in each field of view , positive cell count was graded , 0 - 1 % = 0 , 1 - 10 % = 1 , 10 - 50 % = 2 , 50 - 80 % = 3 , 80 - 100 % = 4 ) . The results are as follows : ( - ) , ihs S1 - 4 are divided into ( + ) , ihs S5 - 8 are divided into ( ++ ) , ihs S9 ~ 12 are divided into ( ++ ) . Ki - 67 results are as follows : brown - yellow particles appear in the cytoplasm of the cells , and the results are determined by immunohistochemistry method . The results are as follows : no staining is 0 , light yellow is 1 , brown - yellow is 2 , brown - brown is 3 - point ;
The percentage of positive cells was scored as percentage of positive cells : 5 visual fields were randomly selected ( x 200 ) , 100 cells were counted in each field of view , the number of positive cells was graded , 0 - 1 % = 0 , 1 - 10 % = 1 , 10 - 50 % = 2 , 50 - 80 % = 3 , 80 - 100 % = 4 ) . The results were as follows : ( - ) , ihs S1 ~ 4 were divided into ( + ) , ihs S5 ~ 8 were divided into ( ++ ) , ihs S9 ~ 12 were divided into ( ++ ) . All the data were treated with SPSS 13.0 statistical software . The count data was analyzed by Pearsonchi - Square Test or Fish ' s exact probability method .
The correlation analysis between the two groups was analyzed by the correlation analysis between the two groups . All the tests were bilateral , P & lt ; 0.05 was statistically significant .

Results :

Expression of FR - 偽 in normal cervical tissue , cervical epithelial neoplasia and cervical cancer

In this experiment , the positive rate of FR - 偽 protein was 6.7 % , the positive rate of FR - 偽 protein was 76.2 % in normal cervical tissue , and the positive rate of FR - 偽 protein in cervical carcinoma was 81.7 % , and the difference was statistically significant ( P < 0.05 ) .

Expression of FR - 偽 protein in different pathological types

There was no significant difference in the expression of FR - 偽 protein in CINI , CINII and CINIII ( P0.05 ) .

Correlation between 3FR - 偽 protein expression and risk factors of cervical cancer

There was no significant difference between the expression of FR - 偽 protein and age in cervical cancer tissues and in cervical epithelial neoplasia . The difference between age 40 and age 鈮

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