【摘要】：腫瘤相關(guān)巨噬細(xì)胞(TAMs)在調(diào)控Ⅰ型子宮內(nèi)膜癌雌激素敏感性中的作用及機(jī)制研究子宮內(nèi)膜癌(Endometrial carcinoma, EC)是女性最常見的生殖系統(tǒng)惡性腫瘤之一,其中70%-80%為Ⅰ型子宮內(nèi)膜樣腺癌,普遍認(rèn)為其發(fā)生與長(zhǎng)期無(wú)孕激素拮抗的雌激素刺激相關(guān)。我們的前期研究發(fā)現(xiàn),Ⅰ型子宮內(nèi)膜癌及其癌前病變患者血清雌激素水平并不高于健康同齡婦女,提示可能存在其他機(jī)制參與Ⅰ型內(nèi)膜癌的發(fā)生發(fā)展。對(duì)其他實(shí)體腫瘤的研究發(fā)現(xiàn),炎癥刺激在腫瘤的發(fā)生發(fā)展中起重要作用。大量流行病學(xué)研究發(fā)現(xiàn),Ⅰ型子宮內(nèi)膜癌與胰島素抵抗密切相關(guān),而慢性炎癥是后者的重要臨床表現(xiàn)之一。也有研究顯示,作為慢性炎癥的具體表現(xiàn)形式,在子宮內(nèi)膜癌病灶存在巨噬細(xì)胞浸潤(rùn),后者與子宮內(nèi)膜癌的不良預(yù)后密切相關(guān)。提示巨噬細(xì)胞浸潤(rùn)可能在Ⅰ型內(nèi)膜癌的發(fā)生發(fā)展中起重要作用。但其作用和機(jī)制還有待進(jìn)一步研究。人體內(nèi)的單核/巨噬細(xì)胞可被誘導(dǎo)活化為不同表型：M1型和M2型,目前,M2型巨噬細(xì)胞被認(rèn)為是在多數(shù)腫瘤中浸潤(rùn)的巨噬細(xì)胞,也被稱為腫瘤相關(guān)巨噬細(xì)胞(Tumor-associated macrophages, TAMs)。以TAMs浸潤(rùn)為主的炎癥反應(yīng)與多種腫瘤的預(yù)后不良有關(guān)。研究發(fā)現(xiàn),巨噬細(xì)胞浸潤(rùn)可調(diào)控腫瘤細(xì)胞激素受體的表達(dá)。在前列腺癌,巨噬細(xì)胞浸潤(rùn)可上調(diào)雄激素受體表達(dá),促進(jìn)雄激素對(duì)前列腺的致癌作用。在乳腺癌,肥胖造成的慢性炎癥狀態(tài)可激活乳腺癌局部的雌激素受體。因此,我們推測(cè)子宮內(nèi)膜病灶局部TAMs浸潤(rùn)可能通過(guò)調(diào)控雌激素受體,增加內(nèi)膜局部對(duì)雌激素的敏感性,促進(jìn)癌癥的發(fā)生和發(fā)展。本研究擬通過(guò)分析巨噬細(xì)胞浸潤(rùn)與子宮內(nèi)膜癌和癌前病變的相關(guān)性,以及巨噬細(xì)胞對(duì)子宮內(nèi)膜癌雌激素受體的調(diào)控及其對(duì)雌激素的敏感性,驗(yàn)證上述假設(shè),并進(jìn)一步分析可能的機(jī)制所在。具體分為四部分：第一部分：Ⅰ型子宮內(nèi)膜癌及內(nèi)膜增生性病變患者血清雌激素水平分析；第二部分：巨噬細(xì)胞浸潤(rùn)與Ⅰ型子宮內(nèi)膜癌及內(nèi)膜增生性病變的相關(guān)性分析；第三部分：TAMs通過(guò)ER α調(diào)控Ⅰ型子宮內(nèi)膜癌細(xì)胞雌激素敏感性；第四部分：TAMs通過(guò)IL-17調(diào)控Ⅰ型子宮內(nèi)膜癌細(xì)胞ER α表達(dá)。第一部分：Ⅰ型子宮內(nèi)膜癌及癌前病變患者血清雌激素水平分析目的：檢測(cè)血清雌激素水平與Ⅰ型子宮內(nèi)膜癌發(fā)生發(fā)展的相關(guān)性。方法：1.收集2011年9月至2013年12月,子宮內(nèi)膜正常及發(fā)生增生性病變患者的一般情況和血清雌二醇(Estradiol,E2)水平資料,包括對(duì)照組39例,增生紊亂69例,單純性增生過(guò)長(zhǎng)168例,復(fù)雜性增生過(guò)長(zhǎng)67例,不典型增生33例,Ⅰ型子宮內(nèi)膜癌25例,共收集病例401例。所有血液學(xué)檢測(cè)均在本院檢驗(yàn)科執(zhí)行。統(tǒng)計(jì)分析血清雌激素水平,采用Graph pad prism 5.0軟件進(jìn)行one-way ANOVA分析,以P=0.05為顯著性統(tǒng)計(jì)學(xué)差異標(biāo)準(zhǔn)。2.收集2011年9月至2013年12月,子宮內(nèi)膜正常及內(nèi)膜癌患者血清,包括對(duì)照組34例,Ⅰ型子宮內(nèi)膜癌34例,共收集病例68例。相同年齡正常及內(nèi)膜癌患者配對(duì)比較。ELISA檢測(cè)血清雌激素水平。采用Graph pad prism 5.0進(jìn)行paired t test分析,以P=0.05為顯著性統(tǒng)計(jì)學(xué)差異標(biāo)準(zhǔn)。結(jié)果：1.臨床資料顯示,雌二醇水平在各組分布分別為：95.00±68.81ng/L、151.8±163.2 ng/L、156.4±178.8 ng/L、138.4±330.0 ng/L、125.5±156.7 ng/L、36.52 ±45.35 ng/L,結(jié)果不具有統(tǒng)計(jì)學(xué)差異(P0.05)。以40歲為界,將患者分為25-40歲和40-55歲兩個(gè)年齡段,25-40歲年齡段中,雌二醇水平在各組分布分別為：107.1 ±70.73 ng/L、140.3±165.2 ng/L、122.1±129.6 ng/L、76.59±59.86 ng/L和120.1±179.2 ng/L；40～55歲年齡段中,雌二醇水平在各組分布分別為：102.2±63.36 ng/L、 161.1±154.5 ng/L、164.0±20.6 ng/L、148.1±152.6 ng/L、148.3±122.5 ng/L和76.86±68.13 ng/L,同一年齡段的各組間雌二醇水平分布不具有統(tǒng)計(jì)學(xué)差異(P0.05)。2.Elisa結(jié)果顯示,雌二醇水平在對(duì)照組與Ⅰ型子宮內(nèi)膜癌組按同一年齡配對(duì)比較：P=0.8338,無(wú)統(tǒng)計(jì)學(xué)意義。結(jié)論：Ⅰ型子宮內(nèi)膜癌及癌前病變患者血清雌激素水平并不高于內(nèi)膜正常婦女。提示可能存在其他機(jī)制參與內(nèi)膜癌的發(fā)生發(fā)展。第二部分：巨噬細(xì)胞浸潤(rùn)與Ⅰ型子宮內(nèi)膜癌及內(nèi)膜增生性病變的相關(guān)性分析目的：檢測(cè)巨噬細(xì)胞在Ⅰ型子宮內(nèi)膜癌及癌前病變中的浸潤(rùn)情況,分析二者間的相關(guān)性。方法：收集正常及增生性病變子宮內(nèi)膜組織共140例：包括增殖期、分泌期、單純型增生、復(fù)雜型增生子宮內(nèi)膜各20例,不典型增生子宮內(nèi)膜、Ⅰ型子宮內(nèi)膜癌各30例,CD68免疫組化染色,計(jì)數(shù)各期別CD68(+)細(xì)胞數(shù),評(píng)價(jià)CD68(+)巨噬細(xì)胞在子宮內(nèi)膜中的浸潤(rùn)情況。采用Graph pad prism 5.0進(jìn)行one-way ANOVA分析,以P=0.05為顯著性統(tǒng)計(jì)學(xué)差異標(biāo)準(zhǔn)。結(jié)果：1.CD68表達(dá)于巨噬細(xì)胞的胞質(zhì)和胞膜。2.各組中CD68+細(xì)胞數(shù)目分別是(每400倍視野)：增殖期：6.641±5.463個(gè)：分泌期：13.82±10.36個(gè)；單純型增生：20.45±10.17個(gè)；復(fù)雜型增生：35.70±14.73個(gè)；不典型增生期：41.98±15.74個(gè)；Ⅰ型子宮內(nèi)膜癌：57.80±25.72個(gè),與Ⅰ型子宮內(nèi)膜癌相比均有P0.001,CD68(+)細(xì)胞數(shù)與子宮內(nèi)膜增生性病變進(jìn)展呈正相關(guān)。3.隨子宮內(nèi)膜增生性病變進(jìn)展,巨噬細(xì)胞浸潤(rùn)部位由血管周圍、間質(zhì)向腺上皮轉(zhuǎn)變。4.隨病變進(jìn)展,巨噬細(xì)胞形態(tài)逐漸成熟。結(jié)論：CD68(+)巨噬細(xì)胞在子宮內(nèi)膜浸潤(rùn)數(shù)目、部位及形態(tài)可能與Ⅰ子宮內(nèi)膜癌發(fā)生發(fā)展有關(guān)。第三部分：TAMs通過(guò)ERα調(diào)控Ⅰ型子宮內(nèi)膜癌細(xì)胞雌激素敏感性目的：研究TAMs在促進(jìn)Ⅰ型子宮內(nèi)膜癌細(xì)胞增殖及內(nèi)膜癌細(xì)胞對(duì)雌二醇敏感性中的作用,并進(jìn)一步分析可能的機(jī)制。方法：采用兩種雌激素受體陽(yáng)性的子宮內(nèi)膜癌細(xì)胞株Ishikawa和HEC-1-A作為研究對(duì)象；培養(yǎng)單核細(xì)胞系THP-1,并誘導(dǎo)為M2型巨噬細(xì)胞,即TAMs;子宮內(nèi)膜癌細(xì)胞與M2型THP-1共培養(yǎng),或經(jīng)M2型THP-1的條件培養(yǎng)基培養(yǎng)后,CCK-8檢測(cè)共培養(yǎng)前后內(nèi)膜癌細(xì)胞增殖情況,以及對(duì)生理劑量(10-9M)E2的敏感性改變,并將此變化與過(guò)表達(dá)ERα或應(yīng)用ER α抑制劑ICI后的趨勢(shì)相比較,采用Graph pad prism 5.0進(jìn)行one-way ANOVA分析,以P=0.05為顯著性統(tǒng)計(jì)學(xué)差異標(biāo)準(zhǔn)；Real-time PCR和Western Blot檢測(cè)內(nèi)膜癌細(xì)胞與M2型THP-1共培養(yǎng)后雌激素受體α (ERα)的表達(dá)變化,并采用細(xì)胞增殖相關(guān)蛋白CyclinDl驗(yàn)證細(xì)胞增殖情況。結(jié)果：1.人M2型THP-1巨噬細(xì)胞誘導(dǎo)活化成功,形態(tài)改變明顯,表型鑒定CD68、CD163、CD204和CD206顯著升高。2.M2型THP-1與Ⅰ型子宮內(nèi)膜癌細(xì)胞Ishikawa或HEC-1-A共培養(yǎng)后,內(nèi)膜癌細(xì)胞增殖均被促進(jìn)；生理劑量的E2對(duì)兩株細(xì)胞的促增殖作用亦被上調(diào)；該現(xiàn)象可被雌激素受體抑制劑ICI抑制。3.M2型THP-1的條件培養(yǎng)基(CM)可時(shí)問(wèn)依賴性促進(jìn)Ishikawa和HEC-1-A細(xì)胞增殖,上調(diào)生理劑量雌激素對(duì)內(nèi)膜癌細(xì)胞的促增殖作用,其促增殖作用同樣可被ICI抑制。4.M2型THP-1的條件培養(yǎng)基(CM)可時(shí)間依賴性上調(diào)雌激素受體α表達(dá),而對(duì)雌激素受體β無(wú)明顯作用；CM可時(shí)間依賴性上調(diào)cyclinDl表達(dá)；5.M2型THP-1對(duì)雌激素促增殖作用的上調(diào)作用與內(nèi)膜癌細(xì)胞過(guò)表達(dá)ER α后細(xì)胞增殖增強(qiáng)情況類似。結(jié)論：TAMs可促進(jìn)子宮內(nèi)膜癌細(xì)胞Ishikawa和HEC-1-A增殖及對(duì)雌激素的敏感性,這種作用可能是通過(guò)上調(diào)內(nèi)膜癌細(xì)胞雌激素受體α實(shí)現(xiàn)的。第四部分：TAMs通過(guò)IL-17調(diào)控Ⅰ型子宮內(nèi)膜癌細(xì)胞ERα表達(dá)目的：篩選TAMs中可能在促進(jìn)內(nèi)膜癌細(xì)胞增殖中起關(guān)鍵作用的炎癥因子,研究其在促Ⅰ型子宮內(nèi)膜癌細(xì)胞增殖及雌激素敏感性方面的作用,并探索可能的機(jī)制。方法：M2型THP-1與子宮內(nèi)膜癌細(xì)胞HEC-1-A經(jīng)Transwell共培養(yǎng)后,提取M2型THP-1細(xì)胞的RNA行Real-time PCR篩選穩(wěn)定作用的炎癥因子；采用CCK-8驗(yàn)證篩選的炎癥因子對(duì)HEC-1-A的促增殖及促雌激素敏感性作用,采用Graph pad prism 5.0進(jìn)行one-way ANOVA分析,以P=0.05為顯著性統(tǒng)計(jì)學(xué)差異標(biāo)準(zhǔn)；Real-time PCR和Western blot驗(yàn)證該炎癥因子對(duì)HEC-1-A的ER α表達(dá)調(diào)控；Western blot驗(yàn)證該炎癥因子和E2共同作用于HEC-1-A細(xì)胞后,對(duì)調(diào)控細(xì)胞增殖的p-AKT信號(hào)通路的活化作用。結(jié)果：1.經(jīng)篩選,與HEC-1-A共培養(yǎng)后M2型THP-1細(xì)胞炎癥因子IL-10和IL-17表達(dá)明顯增加；2.IL-10和IL-17均可上調(diào)E2對(duì)HEC-1-A細(xì)胞的促增殖作用,且IL-17作用更強(qiáng)；3IL-10、IL-17或E2處理后,均可上調(diào)HEC-1-A的ER a表達(dá),且IL-17作用更強(qiáng),進(jìn)一步篩選出IL-17作為研究對(duì)象；4.IL-17與E2共同處理HEC-1-A細(xì)胞后,其p-AKT信號(hào)通路活化顯著增強(qiáng),此作用與HEC-1-A細(xì)胞轉(zhuǎn)染過(guò)表達(dá)ERα后加用E2相似。結(jié)論：TAMs通過(guò)IL-17上調(diào)子宮內(nèi)膜癌細(xì)胞ER a的表達(dá),從而促進(jìn)內(nèi)膜癌細(xì)胞對(duì)雌激素敏感性增加,增強(qiáng)雌激素對(duì)內(nèi)膜癌細(xì)胞的促增殖作用。上述結(jié)果提示：1.血清雌激素水平與Ⅰ型子宮內(nèi)膜增生性病變的進(jìn)展無(wú)顯著相關(guān)性,雌激素對(duì)內(nèi)膜的促癌作用可能存在其他機(jī)制；2.巨噬細(xì)胞浸潤(rùn)與工型子宮內(nèi)膜增生性病變進(jìn)展呈正相關(guān),慢性炎癥可能在Ⅰ型子宮內(nèi)膜癌發(fā)生發(fā)展中發(fā)揮重要作用；3.腫瘤相關(guān)巨噬細(xì)胞通過(guò)上調(diào)內(nèi)膜癌細(xì)胞雌激素受體α表達(dá)增強(qiáng)雌激素對(duì)Ⅰ型子宮內(nèi)膜癌細(xì)胞的促增殖作用,慢性炎癥對(duì)雌激素敏感性的調(diào)控可能是Ⅰ型子宮內(nèi)膜癌發(fā)生發(fā)展的重要原因；4.腫瘤相關(guān)巨噬細(xì)胞通過(guò)分泌炎癥因子,如白介素-17,上調(diào)內(nèi)膜癌細(xì)胞雌激素受體α表達(dá),從而提高內(nèi)膜細(xì)胞對(duì)雌激素敏感性�？赡苁莾�(nèi)膜癌發(fā)生發(fā)展的重要機(jī)制之一本課題研究表明：無(wú)孕激素拮抗的高雌激素水平可能不是雌激素促進(jìn)Ⅰ型子宮內(nèi)膜癌發(fā)生的唯一方式,慢性炎癥調(diào)控的內(nèi)膜雌激素敏感性增加可能為內(nèi)膜癌發(fā)生的機(jī)制提供補(bǔ)充。靶向性抗炎治療可能為預(yù)防Ⅰ型子宮內(nèi)膜癌及癌前病變的發(fā)生提供新的思路。
[Abstract]:The role and mechanism of tumor-associated macrophages (TAMs) in regulating estrogen sensitivity in type I endometrial carcinoma Endometrial carcinoma (EC) is one of the most common female reproductive malignancies, 70% - 80% of which are type I endometrioid adenocarcinoma. It is generally believed that the occurrence of TAMs is associated with estrogen without progesterone antagonism for a long time. Our previous study found that serum estrogen levels in patients with type I endometrial carcinoma and its precancerous lesions were not higher than those in healthy women of the same age, suggesting that there may be other mechanisms involved in the development of type I endometrial carcinoma. A large number of epidemiological studies have found that type I endometrial carcinoma is closely related to insulin resistance, and chronic inflammation is one of the important clinical manifestations of the latter. It is suggested that macrophage infiltration may play an important role in the genesis and development of type I endometrial carcinoma, but its role and mechanism need to be further studied.Monocytes/macrophages in human body can be induced to activate into different phenotypes: M1 and M2. At present, M2 macrophages are considered as infiltrating macrophages in most tumors, also known as macrophages. Tumor-associated macrophages (TAMs). Inflammatory reactions predominantly with TAMs infiltration are associated with poor prognosis in a variety of tumors. Macrophage infiltration can regulate the expression of hormone receptors in tumor cells. In prostate cancer, macrophage infiltration can up-regulate the expression of androgen receptors and promote the effect of androgen on prostate. In breast cancer, chronic inflammation caused by obesity can activate estrogen receptor in breast cancer. Therefore, we speculate that local TAMs infiltration in endometrial lesions may promote the occurrence and development of cancer by regulating estrogen receptor and increasing local sensitivity to estrogen. The relationship between cell infiltration and endometrial carcinoma and precancerous lesions, and the regulation of macrophages on estrogen receptor and their sensitivity to estrogen in endometrial carcinoma were studied to verify the hypothesis and further analyze the possible mechanism. Analysis of serum estrogen level; Part II: Correlation between macrophage infiltration and type I endometrial carcinoma and endometrial hyperplasia; Part III: TAMs regulate estrogen sensitivity of type I endometrial carcinoma cells through ERalpha; Part IV: TAMs regulate ERalpha expression of type I endometrial carcinoma cells through IL-17. Objective: To detect the correlation between serum estrogen levels and the occurrence and development of type I endometrial carcinoma. Methods: 1. To collect the general information and serum estradiol (E2) of patients with normal and proliferative endometrium from September 2011 to December 2013. Data of serum estrogen levels were collected in 401 cases, including 39 cases of control group, 69 cases of hyperplasia disorder, 168 cases of simple hyperplasia, 67 cases of complex hyperplasia, 33 cases of atypical hyperplasia, 25 cases of type I endometrial carcinoma. E-way ANOVA analysis, with P = 0.05 as the significant statistical difference standard. 2. Collected from September 2011 to December 2013, normal endometrial and endometrial cancer patients serum, including 34 cases of control group, 34 cases of type I endometrial cancer, a total of 68 cases. The same age of normal and endometrial cancer patients matched comparison. ELISA detection of serum estrogen levels. Results: 1. The clinical data showed that the distribution of estradiol levels in each group were 95.00 (+ 68.81 ng/L), 151.8 (+ 163.2 ng/L), 156.4 (+ 178.8 ng/L), 138.4 (+ 330.0 ng/L), 125.5 (+ 156.7 ng/L) and 36.52 (+ 45.35 ng/L), respectively. The results were not statistically significant (P 0.05). 05). Patients were divided into 25-40 years old and 40-55 years old. The distribution of estradiol in each group was 107.1 (+ 70.73 ng/L), 140.3 (+ 165.2 ng/L), 122.1 (+ 129.6 ng/L), 76.59 (+ 59.86 ng/L) and 120.1 (+ 179.2 ng/L) in 25-40 years old, respectively. 63.36 ng/L, 161.1+154.5 ng/L, 164.0+20.6 ng/L, 148.1+152.6 ng/L, 148.3+122.5 ng/L and 76.86+68.13 ng/L. There was no significant difference in the distribution of estradiol levels among the same age groups (P 0.05). 2. Elisa results showed that estradiol levels in the control group and the type I endometrial cancer group were matched by the same age: P = 0.8338, no significant difference. Conclusion: The serum estrogen level of patients with type I endometrial carcinoma and precancerous lesions is not higher than that of normal endometrial women, suggesting that there may be other mechanisms involved in the occurrence and development of endometrial carcinoma. Methods: 140 normal and proliferative endometrial tissues were collected, including 20 cases of proliferative endometrium, 20 cases of simple hyperplasia, 20 cases of complex hyperplasia, 30 cases of atypical hyperplasia and 30 cases of type I endometrial carcinoma. The number of CD68 (+) cells in each stage was counted and the infiltration of CD68 (+) macrophages in endometrium was evaluated. Graph pad prism 5.0 was used for one-way ANOVA analysis. P = 0.05 was used as the standard of significant statistical difference. Results: 1. CD68 was expressed in the cytoplasm and membrane of macrophages. 2. The number of CD68 (+) cells in each group was separately analyzed. Proliferative phase: 6.641 + 5.463: Secretory phase: 13.82 + 10.36; Simple hyperplasia: 20.45 + 10.17; Complex hyperplasia: 35.70 + 14.73; Atypical hyperplasia: 41.98 + 15.74; Type I endometrial carcinoma: 57.80 + 25.72; Compared with type I endometrial carcinoma, there were all P 0.001, CD68 (+) cells and sons. With the development of endometrial hyperplasia, the infiltration sites of macrophages changed from perivascular to mesenchymal to glandular epithelial. 4. With the progress of endometrial hyperplasia, the morphology of macrophages gradually matured. Conclusion: The number, location and morphology of CD68 (+) macrophages infiltration in endometrium may be related to the occurrence of endometrial cancer. Part III: TAMs regulate estrogen sensitivity of type I endometrial carcinoma cells through ERalpha. Objective: To study the role of TAMs in promoting the proliferation of type I endometrial carcinoma cells and the sensitivity of endometrial carcinoma cells to estradiol, and to further analyze the possible mechanisms. Membrane cancer cell lines Ishikawa and HEC-1-A were used as research objects; monocyte THP-1 was cultured and induced into M2 macrophages, namely TAMs; endometrial cancer cells were co-cultured with M2 THP-1 or cultured on M2 THP-1 conditioned medium. CCK-8 was used to detect the proliferation of endometrial cancer cells before and after co-culture, and the sensitivity to physiological dose (10-9M) E2. The perceptual changes were compared with those after overexpression of ER-alpha or ICI. One-way ANOVA analysis was performed with Graph pad prism 5.0 and P=0.05 as the significant statistical difference standard. The expression of ER-alpha in endometrial carcinoma cells co-cultured with M2 THP-1 was detected by Real-time PCR and Western Blot. Results: 1. Human M 2 THP-1 macrophages were successfully induced and activated, and the morphological changes were obvious. The phenotypes of CD68, CD163, CD204 and CD206 were significantly increased. 2. M 2 THP-1 co-cultured with Ishikawa or HEC-1-A, the proliferation of endometrial cancer cells was promoted. Physiological dose of E2 also up-regulated the proliferation of the two cell lines; this phenomenon can be inhibited by estrogen receptor inhibitor ICI. 3. M2 THP-1 conditioned medium (CM) can promote the proliferation of Ishikawa and HEC-1-A cells in a time-dependent manner, and up-regulate the proliferation of endometrial cancer cells induced by physiological dose of estrogen, the same as its proliferation-promoting effect. Conditioned medium (CM) inhibited by ICI could up-regulate the expression of estrogen receptor alpha in a time-dependent manner, but had no significant effect on estrogen receptor beta; CM could up-regulate the expression of cyclin Dl in a time-dependent manner; 5. Conclusion: TAMs can promote the proliferation and estrogen sensitivity of endometrial carcinoma cells Ishikawa and HEC-1-A. This effect may be achieved by up-regulating estrogen receptor alpha in endometrial carcinoma cells. Part IV: TAMs regulate the expression of ERalpha in type I endometrial carcinoma cells through IL-17. Objective: To screen TAMs may promote the proliferation of endometrial carcinoma cells. Methods: M2 THP-1 and HEC-1-A cells were co-cultured with Transwell, and RNA of M2 THP-1 cells was extracted to screen stable inflammatory factors by Real-time PCR. Graph pad prism 5.0 was used for one-way ANOVA analysis, and P = 0.05 was used as the significant statistical difference standard; Real-time PCR and Western blot were used to verify the regulation of the inflammatory factors on the expression of ERalpha in HEC-1-A; Western blot was used to verify the inflammation. Results: 1. After co-culture with HEC-1-A, the expression of inflammatory factors IL-10 and IL-17 in M 2 THP-1 cells was significantly increased; 2. Both IL-10 and IL-17 could up-regulate the proliferation of HEC-1-A cells, and the effect of IL-17 was stronger. After treatment with IL-17 or E2, the expression of HEC-1-A was up-regulated, and the effect of IL-17 was stronger. IL-17 was further screened out as the research object. 4. After treatment with IL-17 and E2, the activation of p-AKT signaling pathway in HEC-1-A cells was significantly enhanced, which was similar to that of HEC-1-A cells transfected with ER-a and then treated with E2. These results suggest that: 1. There is no significant correlation between serum estrogen levels and the progression of type I endometrial hyperplasia, and estrogen may have other mechanisms to promote the proliferation of endometrial cancer cells. 2. Macrophage infiltration is positively correlated with the development of type I endometrial hyperplasia. Chronic inflammation may play an important role in the development of type I endometrial carcinoma. 3. Tumor-associated macrophages enhance the proliferation of type I endometrial carcinoma cells by up-regulating the expression of estrogen receptor alpha. The regulation of sex inflammation on estrogen sensitivity may be an important reason for the occurrence and development of type I endometrial carcinoma. This study suggests that high estrogen levels without progesterone antagonism may not be the only way estrogen promotes the development of type I endometrial carcinoma. Increased estrogen sensitivity in the endometrium regulated by chronic inflammation may complement the mechanism of endometrial carcinoma. Targeted anti-inflammatory therapy may prevent type I endometrial carcinoma and New ideas are provided for the occurrence of precancerous lesions.
【學(xué)位授予單位】：復(fù)旦大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R737.33

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