【摘要】：研究背景 子宮肌瘤(uterine fibroids)是育齡婦女生殖器官最常見(jiàn)的良性腫瘤,發(fā)病率呈逐年上升趨勢(shì),臨床發(fā)病率為20%-40%。子宮肌瘤的發(fā)病機(jī)制至今尚不明了。子宮肌瘤是類固醇激素依賴腫瘤,其發(fā)生與遺傳、基因突變等多種因素有關(guān)。異常表達(dá)的基因主要涉及細(xì)胞信號(hào)和傳遞蛋白、離子通道運(yùn)輸?shù)鞍椎�。子宮平滑肌和內(nèi)膜組織中具有豐富的前列腺素合成分泌,通過(guò)自分泌和旁分泌機(jī)制作用于平滑肌細(xì)胞,參與妊娠和非妊娠期的子宮生理效應(yīng)。既往,人們對(duì)子宮肌瘤的發(fā)病機(jī)制的研究中很少涉及前列腺素系統(tǒng)的作用。 前列腺素(Prostaglandins, PG)是多不飽和脂肪酸--花生四烯酸(Arachidonic acid,aa)的衍生物。前列腺素是調(diào)節(jié)子宮收縮的重要激素,而環(huán)氧合酶(Cyclooxygenase-2,COX-2)則是催化aa產(chǎn)生PG途徑當(dāng)中的限速酶,也是非甾體類抗炎藥(Non-steroidal anti-inflammatory drugs,NSAIDs)作用的靶點(diǎn),它不僅調(diào)節(jié)前列腺素的合成,而且廣泛參與腫瘤的發(fā)生發(fā)展過(guò)程。COX-2是COX(Cyclooxygense)的一個(gè)同工酶,一個(gè)即刻早期反應(yīng)基因,通常在大多數(shù)的組織和細(xì)胞中是沒(méi)有表達(dá)的,但是在炎性反應(yīng)、激素和腫瘤啟動(dòng)子中被高度誘導(dǎo),而COX-1(Cyclooxygense-1)的含量并無(wú)明顯的變化。腫瘤組織中COX-2的高度表達(dá),抑制COX-2活性與抗腫瘤作用直接相關(guān),而抗COX-2選擇性抑制劑有抗增殖作用。COX-2衍生的前列腺素E2(prostaglandin E2,PGE2)是一種促生物活性脂質(zhì),是主要的前列腺素。PGE2可促進(jìn)腫瘤生長(zhǎng)。研究表明,在人類多種惡性腫瘤如乳腺癌、腸癌、前列腺癌、肺癌、肝癌、頭頸部腫瘤等,COX-2及PGE2表達(dá)明顯上調(diào),活性增加。鑒于COX-2以及PGE2在腫瘤發(fā)生發(fā)展中的作用,但是目前對(duì)于COX-2和PGE2在子宮肌瘤組織中的表達(dá)及其與子宮肌瘤的發(fā)生發(fā)展的生物學(xué)關(guān)系并不十分清楚。 既往研究表明子宮肌瘤與雌激素和異常細(xì)胞增殖有關(guān),但并不清楚其確切的病理生理機(jī)制。有學(xué)者發(fā)現(xiàn)子宮肌瘤患者的子宮出現(xiàn)異常收縮。最近的研究表明,子宮深層組織,即內(nèi)膜下肌層組織,出現(xiàn)“蠕變收縮”,且平滑肌細(xì)胞運(yùn)動(dòng)機(jī)能下降但擴(kuò)散增強(qiáng)。所有這些研究提示異常的平滑肌收縮可能在子宮肌瘤病理生理過(guò)程中起著重要的作用,而其潛在的分子生物學(xué)機(jī)制尚不清楚,有待進(jìn)一步研究。鈣離子是啟動(dòng)細(xì)胞收縮的重要的上游信號(hào)分子。已知,子宮平滑肌興奮-收縮偶聯(lián)受雙重信號(hào)轉(zhuǎn)導(dǎo)通路的調(diào)節(jié),其中鈣離子信號(hào)轉(zhuǎn)導(dǎo)通路在細(xì)胞應(yīng)激過(guò)程中起著十分重要的作用。鈣通道蛋白是信號(hào)轉(zhuǎn)導(dǎo)過(guò)程中的分子開(kāi)關(guān),他們不僅調(diào)節(jié)細(xì)胞收縮還參與細(xì)胞周期的調(diào)節(jié),這與眾多腫瘤和致癌路徑緊密相關(guān)。瞬時(shí)鈣離子通道蛋白(calcium transient receptor potential channel,TRPC)是介導(dǎo)細(xì)胞Ca2+內(nèi)流的跨膜鈣通道蛋白,不僅調(diào)控細(xì)胞收縮運(yùn)動(dòng),而且參與細(xì)胞增殖周期調(diào)控,與多種腫瘤的發(fā)生發(fā)展關(guān)系密切。近年來(lái),鈣離子通道蛋白的研究領(lǐng)域取得顯著成績(jī)。研究表明,TRPC與異常的子宮收縮有關(guān),但其是否能促進(jìn)子宮肌瘤的增殖尚不清楚。 目前,人們對(duì)存在子宮肌瘤組織中的前列腺素和鈣通道的認(rèn)識(shí)十分有限,一些重要的環(huán)節(jié)需要我們進(jìn)一步求證。我們?cè)诒狙芯恐?擬開(kāi)展COX-2在子宮肌瘤組織及正常平滑肌組織中的表達(dá)及其對(duì)子宮肌瘤平滑肌細(xì)胞的增殖相關(guān)性研究,以明確COX-2與子宮肌瘤發(fā)病的相關(guān)性,并研究子宮肌瘤細(xì)胞鈣離子通道蛋白的表達(dá)情況,及其對(duì)肌瘤細(xì)胞增殖和PGE2分泌的影響、鈣離子的變化。該研究結(jié)果可能對(duì)進(jìn)一步加深我們對(duì)子宮肌瘤的發(fā)生發(fā)展機(jī)制有重要意義。 第一部分子宮肌瘤組織中COX-2的表達(dá)及PGE2的分泌研究 [研究目的] 觀察子宮肌瘤肌組織中COX-2的表達(dá)和PGE2的分泌情況,探討COX-2介導(dǎo)的信號(hào)通路與子宮肌瘤發(fā)病機(jī)制的關(guān)系。 [研究方法] (1)收集30例腹腔鏡下手術(shù)的子宮肌瘤患者的肌瘤組織標(biāo)本和正常瘤旁平滑肌組織標(biāo)本；體外分離和培養(yǎng)子宮肌瘤細(xì)胞和正常瘤旁平滑肌細(xì)胞。 (2)免疫組化、Western blo、RT-PCR法檢測(cè)子宮肌瘤組織和正常瘤旁平滑肌組織中COX-2的表達(dá)。 (3)免疫熒光檢測(cè)觀察COX-2在正常瘤旁平滑肌細(xì)胞和肌瘤細(xì)胞中的表達(dá)。 (4)MTT檢測(cè)子宮肌瘤細(xì)胞和正常瘤旁平滑肌細(xì)胞增殖情況,及COX-2的選擇性抑制劑NS-398和celecoxib對(duì)肌瘤細(xì)胞增殖的影響。 (5)ELISA法定量測(cè)定子宮肌瘤細(xì)胞的PGE2分泌,及NS-398和celecoxib對(duì)肌瘤細(xì)胞PGE2分泌的影響。 [結(jié)果] (1)我們已經(jīng)掌握了人子宮肌瘤平滑肌細(xì)胞及正常人子宮平滑肌細(xì)胞的體外培養(yǎng)方法,建立了成熟及穩(wěn)定的臨床樣本的子宮肌瘤平滑肌細(xì)胞的分離及體外培養(yǎng)技術(shù)。 (2)免疫組化、Western blot實(shí)驗(yàn)及熒光定量PCR實(shí)驗(yàn)：免疫組化的方法檢測(cè)子宮平滑肌組織有少量的COX-2表達(dá),而子宮肌瘤組織表達(dá)顯著增加,表達(dá)部位主要見(jiàn)于細(xì)胞核和細(xì)胞漿。子宮平滑肌中COX-2的陽(yáng)性指數(shù)=11.90,子宮肌瘤中COX-2的陽(yáng)性指數(shù)=46.50,差異有顯著統(tǒng)計(jì)學(xué)意義,*P0.05。Western blot結(jié)果同樣顯示COX-2在子宮肌瘤中的表達(dá)(0.872±0.035)明顯高于子宮平滑肌組織(0.202±0.056),*P0.05。熒光定量PCR顯示COX-2mRNA在子宮肌瘤中的表達(dá)高于正常平滑肌組織,*P0.05。 (3)子宮肌瘤平滑肌細(xì)胞的增殖率約為正常平滑肌細(xì)胞的1.48±0.09倍,組間比較*P0.05。 (4)分別向體外培養(yǎng)的子宮肌瘤平滑肌細(xì)胞及正常子宮組織的平滑肌細(xì)胞添加10μM的NS-398或celecoxib后孵育24小時(shí),加入MTT檢測(cè)細(xì)胞的增殖情況,結(jié)果發(fā)現(xiàn)子宮肌瘤平滑肌細(xì)胞的增殖均受到顯著抑制,但正常子宮平滑肌細(xì)胞增殖則未受到顯著影響。 (5)子宮肌瘤細(xì)胞的PGE2分泌顯著高于正常平滑肌細(xì)胞,*P0.05。 (6)分別向體外培養(yǎng)的子宮肌瘤平滑肌細(xì)胞及正常子宮組織的平滑肌細(xì)胞分別添加10μM的NS-398或celecoxib后孵育24小時(shí),子宮肌瘤平滑肌細(xì)胞的PGE2的分泌均受到顯著抑制,但正常子宮平滑肌細(xì)胞PGE2的分泌則未受到顯著影響。 [結(jié)論及意義] 子宮肌瘤組織中的COX-2顯著高表達(dá),PGE2分泌顯著增加,體外培養(yǎng)子宮肌瘤細(xì)胞增殖速度顯著高于瘤旁正常子宮平滑肌細(xì)胞的增殖速度；而上述現(xiàn)象均可被COX-2的特異性抑制劑顯著抑制。這說(shuō)明肌瘤細(xì)胞COX-2過(guò)表達(dá)與肌瘤細(xì)胞增殖間有相關(guān)性,提示COX-2可能在子宮肌瘤的發(fā)病機(jī)制中起著某種重要作用。 第二部分子宮肌瘤細(xì)胞鈣離子通道蛋白的表達(dá)及其與COX-2誘導(dǎo)的PGE2分泌的關(guān)系研究 [研究目的] 觀察比較子宮肌瘤細(xì)胞與子宮平滑肌細(xì)胞的鈣離子水平濃度和鈣離子通道蛋白的表達(dá)情況,采用分別針對(duì)TRPC1及TRPM7的shRNA慢病毒載體干擾相應(yīng)蛋白表達(dá)后,觀察子宮肌瘤細(xì)胞的增殖、PGE2的分泌及鈣離子的變化,從而鑒定鈣離子信號(hào)轉(zhuǎn)導(dǎo)通路與子宮肌瘤間的關(guān)系。 [研究方法] (1)收集30例腹腔鏡下手術(shù)的子宮肌瘤患者的子宮肌瘤組織和相應(yīng)的瘤旁正常平滑肌組織,子宮肌瘤細(xì)胞和正常瘤旁平滑肌細(xì)胞的分離及體外培養(yǎng)。 (2)激光掃描共聚焦顯微鏡檢測(cè)細(xì)胞內(nèi)游離鈣離子濃度。 (3) RT-PCR及Western blot法檢測(cè)子宮肌瘤組織和子宮平滑肌組織中鈣通道亞型的表達(dá)。 (4)采用攜帶有TRPC1shRNA和TRPM7shRNA的GIPZ慢病毒載體行RNA干預(yù)實(shí)驗(yàn)。 (5)MTT檢測(cè)RNA干擾TRPC1或TRPM7表達(dá)后對(duì)肌瘤細(xì)胞增殖的影響。 (6) ELISA檢測(cè)RNA干擾TRPC1或TRPM7表達(dá)后對(duì)子宮肌瘤細(xì)胞分泌PGE2的影響。 (7)激光掃描共聚焦顯微鏡檢測(cè)RNA干擾TRPC1和TRPM7后鈣離子的變化。 [結(jié)果] (1)激光共聚焦顯微鏡檢測(cè)胞內(nèi)游離鈣離子濃度顯示,肌瘤組的鈣濃度值25.443土3.203(n=30)顯著高于癌旁正常平滑肌組13.223±1.450(n=30),*P0.05。 (2)PCR定量方法檢測(cè)并比較子宮肌瘤組織和瘤旁正常平滑肌組織間的鈣通道亞型mRNA表達(dá)顯示,子宮肌瘤組的TRPM7和TRPC1表達(dá)顯著高于正常瘤旁平滑肌組,組間比較*P0.05,而兩組間鈣通道的其他亞型的表達(dá)無(wú)顯著差異,P0.05。免疫印跡分析檢測(cè)并比較子宮肌瘤組織和瘤旁正常平滑肌組織間的鈣通道亞型mRNA表達(dá)顯示,子宮肌瘤組的TRPM7和TRPC1表達(dá)顯著高于正常瘤旁平滑肌組,組間比較*P0.05,而兩組間鈣通道的其他亞型的表達(dá)無(wú)顯著性差異,P0.05。 (3)采用shRNA病毒顯著降低肌瘤細(xì)胞TRPC1的表達(dá)后,發(fā)現(xiàn)子宮肌瘤細(xì)胞增殖受到顯著抑制,且子宮肌瘤細(xì)胞PGE2的分泌受到顯著抑制,鈣離子水平降低,*P0.05。 (4)采用shRNA病毒顯著降低肌瘤細(xì)胞TRPM7的表達(dá)后,發(fā)現(xiàn)子宮肌瘤細(xì)胞增殖受到顯著抑制,且子宮肌瘤細(xì)胞的PGE2的分泌受到顯著抑制,鈣離子水平降低,*P0.05。 [結(jié)論及意義] 子宮肌瘤細(xì)胞內(nèi)的游離鈣離子濃度顯著較高；子宮肌瘤組織的鈣離子通道蛋白TRPC1和TRPM7顯著高表達(dá)；采用RNA干擾技術(shù)沉默TRPC1和TRPM7表達(dá)后,子宮肌瘤細(xì)胞增殖明顯受抑制,PGE2的產(chǎn)生分泌明顯降低,并且鈣離子水平下降。 本研究的創(chuàng)新點(diǎn) 1.我們從前列腺素代謝入手,研究COX2、PGE2在子宮肌瘤發(fā)生發(fā)展中的作用,為子宮肌瘤的發(fā)病機(jī)制研究提供新思路。 2.從NSAIDs著手,試圖從抑制劑阻斷子宮肌瘤的發(fā)展,為子宮肌瘤的防治提供新思路。 3.本研究發(fā)現(xiàn)了子宮肌瘤細(xì)胞的鈣離子通道蛋白TRPC1和TRPM7表達(dá)顯著增高,并通過(guò)RNA干擾技術(shù)分別將兩種基因沉默后發(fā)現(xiàn),肌瘤細(xì)胞的增殖均受到顯著抑制,其PGE2分泌亦均受到顯著抑制,并且鈣離子水平下降。這說(shuō)明TRPC1和TRPM7可能參與了子宮肌瘤發(fā)生發(fā)展過(guò)程,而兩者所介導(dǎo)的鈣離子的轉(zhuǎn)運(yùn)路徑可能與COX2誘導(dǎo)的PGE2的分泌路徑存在信號(hào)交互作用。
[Abstract]:Research background
Uterine fibroids are the most common benign tumors in reproductive organs of women of childbearing age. The incidence of uterine fibroids is increasing year by year. The clinical incidence is 20%-40%. The pathogenesis of uterine fibroids is still unknown. Uterine fibroids are steroid-dependent tumors, which are related to many factors, such as heredity, gene mutation and so on. In the past, the pathogenesis of uterine leiomyoma has been studied in the past. The role of prostaglandin system is rarely involved in the study.
Prostaglandins (PG) are derivatives of polyunsaturated fatty acids, arachidonic acid (aa). Prostaglandins are important hormones that regulate uterine contraction, while cyclooxygenase-2 (COX-2) is the rate-limiting enzyme in the pathway of PG production by AA and is also a non-steroidal anti-inflammation drug. COX-2 is an isoenzyme of COX (Cyclooxygense), an immediate early-response gene that is not normally expressed in most tissues and cells, but is involved in inflammatory reactions, hormones, and tumor promoters. High expression of COX-2 and inhibition of COX-2 activity are directly related to the anti-tumor effect, while anti-COX-2 selective inhibitors have anti-proliferative effect. COX-2-derived prostaglandin E2 (PGE2) is a bioactive lipid and is the main one. Prostaglandin-PGE2 promotes tumor growth. Studies have shown that COX-2 and PGE2 are up-regulated and their activities are increased in many human malignant tumors such as breast, bowel, prostate, lung, liver, head and neck tumors. The expression and its biological relationship with the occurrence and development of uterine leiomyoma are not very clear.
Previous studies have shown that uterine leiomyoma is associated with estrogen and abnormal cell proliferation, but the exact pathophysiological mechanism is unclear. Some scholars have found abnormal contraction of the uterus in patients with uterine leiomyoma. Recent studies have shown that "creep contraction" occurs in the deep uterine tissue, i.e. the subendometrial myometrium, and smooth muscle cell motility. All these studies suggest that abnormal smooth muscle contraction may play an important role in the pathophysiological process of uterine leiomyoma, and the underlying molecular biological mechanisms remain unclear and need further study. Calcium ion is an important upstream signaling molecule that initiates cell contraction. Condensation coupling is regulated by dual signal transduction pathways, in which calcium ion signal transduction pathways play an important role in cell stress. Calcium channel proteins are molecular switches in signal transduction. They not only regulate cell contraction but also participate in cell cycle regulation, which is closely related to many tumors and carcinogenic pathways. Calcium transient receptor potential channel (TRPC) is a transmembrane calcium channel protein that mediates the influx of Ca2+ into cells. It not only regulates cell contraction, but also participates in the regulation of cell proliferation cycle. TRPC is closely related to the occurrence and development of various tumors. Studies have shown that TRPC is associated with abnormal uterine contractions, but whether TRPC can promote the proliferation of uterine leiomyomas remains unclear.
At present, the understanding of prostaglandins and calcium channels in uterine leiomyoma is very limited, and some important links need further confirmation. In this study, we intend to study the expression of COX-2 in uterine leiomyoma tissue and normal smooth muscle tissue and its correlation with the proliferation of uterine leiomyoma smooth muscle cells. To clarify the relationship between COX-2 and the pathogenesis of uterine leiomyoma, and to study the expression of calcium channel protein in uterine leiomyoma cells, its effect on the proliferation of leiomyoma cells, the secretion of PGE2, and the changes of calcium ions.
Part one COX-2 expression and PGE2 secretion in uterine fibroids
[research purposes]
To observe the expression of COX-2 and the secretion of PGE2 in uterine leiomyoma, and to explore the relationship between COX-2-mediated signaling pathway and the pathogenesis of uterine leiomyoma.
[research methods]
(1) Samples of leiomyoma tissue and normal paraneoplastic smooth muscle tissue were collected from 30 patients with uterine leiomyoma undergoing laparoscopic surgery, and myoma cells and normal paraneoplastic smooth muscle cells were isolated and cultured in vitro.
(2) Immunohistochemistry, Western Blo and RT-PCR were used to detect the expression of COX-2 in uterine leiomyoma tissues and normal paraneoplastic smooth muscle tissues.
(3) immunofluorescence assay was used to observe the expression of COX-2 in normal tumor adjacent smooth muscle cells and myoma cells.
(4) MTT assay was used to detect the proliferation of uterine leiomyoma cells and normal paraneoplastic smooth muscle cells, and the effects of COX-2 selective inhibitors NS-398 and celecoxib on the proliferation of leiomyoma cells.
(5) Quantitative determination of PGE2 secretion by ELISA and the effects of NS-398 and celecoxib on PGE2 secretion by hysteromyoma cells.
[results]
(1) We have mastered the culture methods of human leiomyoma smooth muscle cells and normal human leiomyoma smooth muscle cells in vitro, and established the isolation and culture techniques of mature and stable clinical samples of leiomyoma smooth muscle cells in vitro.
(2) Immunohistochemistry, Western blot and fluorescence quantitative PCR: Immunohistochemistry showed that there was a small amount of COX-2 expression in uterine leiomyoma tissues, but the expression of COX-2 in uterine leiomyoma tissues was significantly increased, mainly in the nucleus and cytoplasm. The positive index of COX-2 in uterine leiomyoma was 11.90, and the positive index of COX-2 in uterine leiomyoma tissues was 11.90. The expression of COX-2 in uterine leiomyoma was significantly higher than that in uterine leiomyoma (0.872 (+ 0.035)), * P 0.05. The expression of COX-2 mRNA in uterine leiomyoma was higher than that in normal uterine leiomyoma, * P 0.05.
(3) The proliferation rate of leiomyocytes in uterine leiomyoma was 1.48 65507
(4) The proliferation of uterine leiomyoma smooth muscle cells and normal uterine smooth muscle cells was detected by MTT after incubation for 24 hours. The results showed that the proliferation of leiomyoma smooth muscle cells was significantly inhibited, but the proliferation of normal uterine smooth muscle cells was not. Significant impact.
(5) PGE2 secretion of uterine leiomyoma cells was significantly higher than that of normal smooth muscle cells, *P0.05.
(6) The secretion of PGE2 in uterine leiomyoma smooth muscle cells was significantly inhibited 24 hours after incubation with NS-398 or celecoxib of 10 mu M respectively, but the secretion of PGE2 in normal uterine smooth muscle cells was not significantly affected.
[Conclusion and significance]
The expression of COX-2 and the secretion of PGE2 were significantly increased in uterine leiomyoma tissues, and the proliferation rate of cultured uterine leiomyoma cells was significantly higher than that of normal uterine leiomyoma cells in vitro. Correlation suggests that COX-2 may play an important role in the pathogenesis of uterine leiomyoma.
Part 2 Expression of calcium channel protein in uterine leiomyoma cells and its relationship with COX-2-induced PGE2 secretion
[research purposes]
To observe and compare the level of calcium ion and the expression of calcium channel protein in uterine leiomyoma cells and uterine smooth muscle cells, the proliferation of uterine leiomyoma cells, the secretion of PGE2 and the change of calcium ion were observed after the expression of corresponding protein was interfered by shRNA lentiviral vector targeting TRPC1 and TRPM7 respectively, and the calcium ion signal transduction was identified. The relationship between conduction pathways and uterine fibroids.
[research methods]
(1) Isolation and culture of uterine leiomyoma tissues and normal paraneoplastic smooth muscle tissues, uterine leiomyoma cells and normal paraneoplastic smooth muscle cells from 30 patients with uterine leiomyoma undergoing laparoscopic surgery.
(2) intracellular free calcium concentration was detected by laser scanning confocal microscope.
(3) The expression of calcium channel subtypes was detected by RT-PCR and Western blot.
(4) TRPM7shRNA lentiviral vectors carrying TRPC1shRNA and GIPZ were used for RNA intervention experiment.
(5) MTT detected the effect of RNA interference on the proliferation of myoma cells after the expression of TRPC1 or TRPM7.
(6) ELISA can detect the effect of RNA interference on TRPC1 and TRPM7 expression in the secretion of PGE2 from uterine leiomyoma cells.
(7) laser scanning confocal microscopy was used to detect the changes of calcium ions after RNA interferes with TRPC1 and TRPM7.
[results]
(1) Laser confocal microscopy showed that the intracellular free calcium concentration in leiomyoma group was 25.443 soil 3.203 (n=30), which was significantly higher than that in normal smooth muscle group (13.223 + 1.450) (n = 30), * P 0.05.
(2) The expression of TRPM7 and TRPC1 mRNA in uterine leiomyoma tissues and normal smooth muscle tissues adjacent to uterine leiomyoma were detected and compared by PCR quantitative method. The expression of TRPM7 and TRPC1 mRNA in uterine leiomyoma group was significantly higher than that in normal smooth muscle tissues adjacent to uterine leiomyoma group, and the expression of other subtypes of calcium channel between the two groups was not significantly different, P 0.05. The expression of TRPM7 and TRPC1 was significantly higher in uterine leiomyoma group than in normal paraneoplastic leiomyoma group (P 0.05). There was no significant difference in the expression of other subtypes of calcium channel between the two groups (P 0.05).
(3) After the expression of TRPC1 was significantly decreased by shRNA virus, the proliferation of uterine leiomyoma cells was significantly inhibited, the secretion of PGE2 was significantly inhibited, and the level of calcium was decreased, * P 0.05.
(4) After the expression of TRPM7 was significantly decreased by shRNA virus, the proliferation of uterine leiomyoma cells was significantly inhibited, the secretion of PGE2 was significantly inhibited, and the level of calcium was decreased, * P 0.05.
[Conclusion and significance]
The free calcium concentration in uterine leiomyoma cells was significantly higher; the expression of calcium channel proteins TRPC1 and TRPM7 was significantly higher in uterine leiomyoma tissues; the proliferation of uterine leiomyoma cells was significantly inhibited, the production and secretion of PGE2 were significantly decreased, and the level of calcium ion was significantly decreased after TRPC1 and TRPM7 were silenced by RNA interference.
The innovation of this research
1. Starting with prostaglandin metabolism, we studied the role of COX2 and PGE2 in the occurrence and development of uterine leiomyoma, and provided new ideas for the pathogenesis of uterine leiomyoma.
2. Starting with NSAIDs, we try to block the development of uterine fibroids from inhibitors, and provide new ideas for the prevention and treatment of uterine fibroids.
3. This study found that the expression of calcium channel protein TRPC1 and TRPM7 in uterine leiomyoma cells was significantly increased, and the proliferation of leiomyoma cells was significantly inhibited, the secretion of PGE2 was significantly inhibited, and the level of calcium was decreased by RNA interference. The development of uterine leiomyoma may be mediated by calcium transport pathway and COX2-induced PGE2 secretion pathway.
【學(xué)位授予單位】：山東大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2014
【分類號(hào)】：R737.33

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