本文選題：PRR11 + 胃癌�。� 參考：《第三軍醫(yī)大學(xué)》2015年博士論文

【摘要】：研究背景和目的胃癌(Gastric cancer,GC)是世界范圍內(nèi)最常見的惡性腫瘤之一,其發(fā)病率居全部惡性腫瘤的第四位。2008年全球胃癌患者新發(fā)病例約100萬例,當(dāng)年因胃癌死亡的患者大約為73萬例,約占全部惡性腫瘤總發(fā)病的8%,及總死亡的10%。其中,超過70%的新發(fā)胃癌患者和死亡患者出現(xiàn)在發(fā)展中國家,主要分布在東歐、南美洲及亞洲地區(qū)。即使在過去數(shù)十年里,胃癌的診斷和治療已經(jīng)取得了巨大的進展,患者的死亡有所改善,但在全世界范圍內(nèi),其仍然是惡性腫瘤相關(guān)死亡的第二大原因。造成這種情況的主要原因是大多數(shù)胃癌患者在診斷時已處于胃癌晚期,失去手術(shù)根治的機會,得不到有效的治療。目前,已經(jīng)報道了幾個新的影響胃癌患者預(yù)后和/或治療效果的組織蛋白標(biāo)志物,然而,僅有少量的蛋白標(biāo)志物被用于臨床實踐。因此,發(fā)現(xiàn)可精確預(yù)測胃癌患者預(yù)后或預(yù)測治療效果的新分子標(biāo)志物,對其進行深入研究,制備出相應(yīng)的分子靶向藥物,對于胃癌患者來說是非常迫切的需要。富含脯氨酸蛋白11(Proline-rich protein 11,PRR11)基因是一個新近鑒定的具有癌基因潛能基因。Ji Ying等于2013年初步研究了其在肺癌中的作用,并揭示其在肺癌細(xì)胞周期和肺癌形成中具有關(guān)鍵性的作用,可能在肺癌中作為一個新的潛在的診斷和/或治療靶點。目前,PRR11基因在胃癌中的表達模式及其對胃癌影響的認(rèn)識尚未見報道,需要進行研究,可能為胃癌的診斷與治療提供新的理論思路和臨床實踐。在這個研究中,我們收集了216例胃癌患者的胃癌組織及其正常胃粘膜組織進行了PRR11蛋白表達狀態(tài)評估,分析了PRR11蛋白表達與胃癌臨床病理參數(shù)之間的關(guān)系,判斷PRR11蛋白表達是否可以預(yù)測胃癌患者的預(yù)后及其與哪些胃癌臨床病例參數(shù)相關(guān)。在PRR11蛋白高表達的胃癌細(xì)胞中沉默PRR11基因后,分析其對胃癌細(xì)胞增殖、克隆形成及裸鼠體內(nèi)移植瘤生長的影響�；虮磉_譜分析PRR11基因沉默后引起一些基因表達變化顯著,篩選表達變化顯著的基因做相關(guān)性分析。我們篩選出表達顯著降低的基因三螺旋重復(fù)膠原蛋白1(Collagen triple helix repeat containing 1,CTHRC1)及表達顯著增加的基因羧肽酶A抑制因子(Latexin,LXN),免疫組化檢測PRR11與CTHRC1及LXN共表達情況,分析PRR11與CTHRC1及LXN表達的相關(guān)性。研究方法1.組織樣本及患者信息從病理資料庫中選擇符合以下條件的胃癌患者216例:2001年1月至2005年12月期間,在解放軍上海長海醫(yī)院普通外科進行手術(shù)切除治療的胃癌患者,隨訪日期至2010年12月。并從組織標(biāo)本庫中挑出HE染色切片及組織蠟塊,經(jīng)病理科兩位經(jīng)驗豐富的病例專家獨立確定為胃癌。胃癌患者臨床病理參數(shù)包括年齡、性別、腫瘤大小、T分期、N分期、TNM分期及腫瘤分化程度。本研究所有組織樣本的獲得,均得到患者的同意,并簽訂知情同意書,并得到長海醫(yī)院倫理委員會許可。2.組織芯片(Tissue microarray,TMA)及免疫組化(Immunohistochemistry,IHC)由兩位經(jīng)驗豐富的病理學(xué)專業(yè)人員對所有患者的HE染色組織玻片均進行了會診及鑒定,確定為胃癌組織,并在具有代表性的部位進行標(biāo)記。從每個供體蠟塊上打孔取出直徑為1.5mm的組織柱,放入受體石蠟中。4um厚的切片被放到包被3-氨基丙基三乙氧基硅烷(3-aminopropyltriethoxysilane,APTS)的玻片上。抗體稀釋如下:抗-PRR11(1:100稀釋),抗-LXN(1:100稀釋);抗-CTHRC1(1:100稀釋)。鏈霉親和素—過氧化物酶法(Streptavidin-perosidase,S-P法)免疫組化染色使抗體結(jié)合組織顯色,蘇木素復(fù)染。PRR11、LXN、CTHRC1免疫組化染色由兩個獨立的專家在奧林帕斯CX31顯微鏡下觀察。腫瘤細(xì)胞PRR11蛋白染色(胞漿呈棕色)數(shù)量10%即為陽性。3.細(xì)胞系及培養(yǎng)條件在5個GC細(xì)胞系(SGC7901,MKN45,MKN28,HGC27,MGC803)中發(fā)現(xiàn)PRR11蛋白均有表達。GC細(xì)胞系培養(yǎng)于杜爾伯克改良伊格爾培養(yǎng)基(Dulbecco modified Eagle medium,DMEM)加上10%的胎牛血清(Fetal calf serum,FBS)中,并置于37℃,含5%CO2的培養(yǎng)箱中培養(yǎng),細(xì)胞覆蓋率達80%左右傳代培養(yǎng)。4.通過載有sh RNA的慢病毒敲低胃癌細(xì)胞中的PRR11蛋白表達包含靶向PRR11的短發(fā)夾RNA(Short hairpin RNA,sh RNA)(5’-ACG CAG GCC UUA AGG AGA ATT-3’)的慢病毒由GENECHEM公司構(gòu)建,含6ug/ml聚凝胺,用來轉(zhuǎn)染胃癌細(xì)胞。轉(zhuǎn)染后的胃癌細(xì)胞用嘌呤霉素進行選擇,利用蛋白質(zhì)免疫印跡(Immunoblotting,Western blot)方法檢測PRR11敲低的效果。5.細(xì)胞增殖分析和克隆形式分析以每孔5000個細(xì)胞的密度,將穩(wěn)定轉(zhuǎn)染的PRR11-sh RNA細(xì)胞(PRR11-KO細(xì)胞)或空載體陰性對照細(xì)胞(PRR11-NC細(xì)胞)接種于96孔板中,各設(shè)三個副孔,利用細(xì)胞計數(shù)盒(Cell counting Kit-8,CCK8),通過計算24小時及48小時的吸光度,檢測細(xì)胞增殖率。同上,以每孔500個細(xì)胞的密度,將PRR11-KO細(xì)胞和PRR11-NC細(xì)胞接種于6孔板中,各三孔。孵育兩周后,甲醇/丙酮(1:1)固定,結(jié)晶紫染色,計算超過50個細(xì)胞的克隆數(shù)量。6. Western blot分別準(zhǔn)備5種胃癌細(xì)胞系總蛋白,利用兔抗人PRR11、CTHRC1、LNX的多克隆抗體(1:1000稀釋),及辣根過氧化物酶(Horse radish peroxidase,HRP)結(jié)合的山羊抗兔Ig G抗體(1:10000稀釋)作為二抗,β-actin作為內(nèi)參。根據(jù)生產(chǎn)廠家的說明,利用Amersham增強化學(xué)發(fā)光系統(tǒng)進行蛋白顯影及灰度值檢測。7.實時定量逆轉(zhuǎn)錄PCR(q RT-PCR)分析根據(jù)說明,利用SYBR1 Premix Ex Taq TM試劑盒進行實時定量逆轉(zhuǎn)錄聚合酶鏈?zhǔn)椒磻?yīng)(Quantitative Real-time reverse transcription PCR,q RT-PCR),甘油醛-3-磷酸脫氫酶(Glyceraldehyde-3-phosphate dehydrogenase,GAPDH)作為內(nèi)參。PCR程序為95℃1分鐘,然后在95℃15秒,60℃31秒,共40個循環(huán)。利用ΔCt方法計算倍增表達。利用ABI PRISM 7300系統(tǒng)進行PCR檢測及分析。引物利用如下:PRR11:前5’-CGT ATC TGC CAC CGA GAA CTT-3’,反:5’-GAG ATG GTC TTC AGT GCT TCC T-3’;GAPDH:前5’-TGA CTT CAA CAG CGA CAC CCA-3’,反:5’-CAC CCT GTT GCT GTA GCC AAA-3。8.動物模型通過裸鼠皮下細(xì)胞移植瘤模型評估PRR11敲低后的SGC7901細(xì)胞即PRR11-KO細(xì)胞體內(nèi)成瘤及生長能力變化。PRR11-KO細(xì)胞和PRR11-NC細(xì)胞(1×106個細(xì)胞0.1ml PBS)分別注射于4周齡雌性Balb/c裸鼠右側(cè)腋下。每三天測量腫瘤直徑。移植后2周,處死所有裸鼠。收集腫瘤并測量腫瘤體積,V=0.52(長×寬×高)。9.統(tǒng)計分析統(tǒng)計分析利用SPSS 13.0軟件。PRR11蛋白表達和胃癌臨床病理參數(shù)之間的關(guān)系分析采用χ2檢驗(chi-squared分布)。根據(jù)PRR11蛋白表達情況對胃癌患者分層,利用Kaplan-Meier方法進行生存分析。單因素和多因素分析基于Cox比例風(fēng)險回歸模型。實驗計量資料以平均值±標(biāo)準(zhǔn)差的方式呈現(xiàn),計量資料分析采用Student’t-檢驗。p㩳0.05認(rèn)為有統(tǒng)計學(xué)意義。研究結(jié)果1.胃癌組織及正常胃粘膜組織中PRR11蛋白表達差異顯著,PRR11蛋白表達與胃癌臨床病理參數(shù)相關(guān)我們對216例確診為胃癌的患者樣本進行免疫組化評估。結(jié)果顯示PRR11蛋白在正常胃粘膜組織中的免疫染色為低強度,而在部分胃癌組織中的免疫染色為高強度。在216例胃癌組織樣本中有107例(49.5%)PRR11蛋白表達陽性,109例(50.5%)PRR11蛋白表達陰性。q RT-PCR及Western blot分析進一步證明,相對于相應(yīng)的正常胃粘膜組織,PRR11 m RNA及蛋白在胃癌組織樣本中高表達。另外在SGC7901,MKN45,MKN28,HGC27和MGC803等5個胃癌細(xì)胞系中,我們發(fā)現(xiàn)PRR11蛋白均有表達。PRR11蛋白表達水平與多個胃癌臨床病理參數(shù)之間關(guān)系的分析顯示:PRR11蛋白表達與患者年齡、性別、腫瘤大小、N分期無明顯相關(guān)性,而與T分期、腫瘤分化程度、TNM分期顯著相關(guān)。該結(jié)果提示PRR11蛋白高表達可能與胃癌的侵襲及轉(zhuǎn)移相關(guān)。2.PRR11表達上調(diào)和胃癌患者生存降低相關(guān)216位胃癌患者中,122例在隨訪期間死亡,中位生存期(Median survival time,MST)為51.5個月。PRR11蛋白表達陰性的胃癌患者中位生存期為74.5個月,而PRR11蛋白表達陽性的胃癌患者中位生存期為46.4個月。另外,單變量COX回歸分析顯示腫瘤大小、腫瘤T分期、區(qū)域淋巴結(jié)轉(zhuǎn)移、TNM分期、腫瘤分化程度及PRR11表達與患者總生存(Overall survival,OS)顯著相關(guān)。多變量分析顯示腫瘤大小、腫瘤分化程度、PRR11表達是胃癌患者獨立的預(yù)后因素。把患者分成I/II期和III/IV期兩個亞群,分析PRR11表達對各亞群患者生存時間的影響,結(jié)果顯示在I/II期與III/IV期兩個亞群中,相對于PRR11不表達患者,PRR11高表達患者預(yù)后較差(P0.05)。3.PRR11低表達與SGC7901胃癌細(xì)胞體外增殖、克隆形成減少以及體內(nèi)腫瘤生長緩慢相關(guān)利用慢病毒載荷PRR11 sh RNA穩(wěn)定沉默SGC7901細(xì)胞系中PRR11表達。Western blot方法檢測確證PRR11低表達SGC7901細(xì)胞系(PRR11-KO)成功建立,PRR11-KO細(xì)胞中PRR11表達顯著降低,PRR11低表達與SGC7901細(xì)胞系體外增殖及克隆形成明顯下降相關(guān)。同樣,PRR11表達下調(diào)導(dǎo)致SGC7901細(xì)胞系裸鼠體內(nèi)生長延緩。4.PRR11表達敲低后導(dǎo)致CTHRC1表達下調(diào)及LXN表達上調(diào)我們利用基因表達譜進行差異表達分析發(fā)現(xiàn),PRR11表達下降導(dǎo)致CTHRC1m RNA表達下調(diào)及LXN m RNA表達上調(diào)。進一步利用CTHRC1蛋白和LXN蛋白抗體進行了Western blot分析,來探索這些蛋白是否隨著PRR11表達降低而改變。結(jié)果顯示:與PRR11-NC細(xì)胞相比較,在PRR11-KO細(xì)胞中CTHRC1蛋白表達下調(diào),LXN蛋白表達上調(diào)。通過胃癌組織免疫組化檢測進行共表達分析。數(shù)據(jù)顯示胃癌組織中CTHRC1蛋白表達與PRR11蛋白表達正相關(guān)(r=0.299,p0.001),LNX蛋白表達和PRR11蛋白表達負(fù)相關(guān)(r=-0.188,p=0.005)。而CTHRC1蛋白表達和LXN蛋白表達無相關(guān)性(r=-0.042,p=0.539)。結(jié)論1.PRR11蛋白在胃癌組織中高表達,在正常胃粘膜組織中低表達,且與胃癌分化程度、T分期、TNM分期等臨床病理參數(shù)相關(guān)。這提示PRR11蛋白高表達可能與胃癌細(xì)胞的侵襲與轉(zhuǎn)移相關(guān)。2.多因素分析顯示胃癌腫塊大小、分化程度與PRR11蛋白表達是胃癌患者總生存的獨立預(yù)后因素。3.PRR11蛋白在胃癌細(xì)胞系中表達,PRR11蛋白低表達降低了胃癌細(xì)胞體外增殖、克隆形成能力及體內(nèi)腫瘤生長能力。4.PRR11表達敲低后引起胃癌細(xì)胞中CTHRC1表達下調(diào)以及LXN表達上調(diào)。5.進一步研究PRR11蛋白對胃癌發(fā)生、發(fā)展的作用機制,可為臨床防治胃癌提供新的作用靶點。
[Abstract]:Background and objective Gastric cancer (GC) is one of the most common malignant tumors in the world, with the incidence of fourth new cases of global cancer in the world, about 1 million cases of global gastric cancer, and about 730 thousand patients died of gastric cancer in those years, accounting for 8% of the total incidence of all malignant tumors and the 10%. of total death. Among them, more than 70% of new and dead patients were found in developing countries, mainly in Eastern Europe, South America and Asia. Even in the past few decades, the diagnosis and treatment of gastric cancer had made great progress and the death of the patients improved, but in the world, it was still associated with the death of malignant tumors. Second major reasons. The main reason for this is that most gastric cancer patients are in the advanced stage of gastric cancer at the time of diagnosis, lose the chance of radical operation, and have no effective treatment. At present, several new egg white markers have been reported to affect the prognosis and / or treatment effect of gastric cancer patients. However, only a small amount of protein markers are used. It is used in clinical practice. Therefore, it is very urgent to find a new molecular marker to predict the prognosis of gastric cancer and predict the therapeutic effect of gastric cancer accurately. It is very urgent for the patients with gastric cancer to prepare the corresponding molecular targeted drugs. The gene of proline protein 11 (Proline-rich protein 11, PRR11) is a new approach. The identification of the oncogene potential gene.Ji Ying is equal to the preliminary study of its role in lung cancer in 2013, and reveals its key role in the cell cycle of lung cancer and the formation of lung cancer. It may be a new potential diagnostic and / or therapeutic target in lung cancer. At present, the expression pattern and its expression of PRR11 gene in gastric cancer The understanding of the influence of gastric cancer has not yet been reported. It needs to be studied, which may provide new theoretical ideas and clinical practice for the diagnosis and treatment of gastric cancer. In this study, we collected 216 cases of gastric cancer and normal gastric mucosa to evaluate the expression of PRR11 protein, and analyzed the expression of PRR11 protein and gastric cancer. The relationship between the clinicopathological parameters to determine whether the expression of PRR11 protein can predict the prognosis of gastric cancer patients and which of the clinical case parameters of gastric cancer. After the silencing of PRR11 gene in the gastric cancer cells with high expression of PRR11 protein, the effects of the gene expression on the proliferation of gastric cancer cells, clonogenic formation and the growth of xenografts in nude mice. A significant change in gene expression was caused by PRR11 gene silencing, and a significant gene was screened for correlation analysis. We screened three spiral repeat collagen 1 (Collagen triple helix repeat containing 1, CTHRC1) and a significant increase in expression of the gene carboxypeptidase A inhibitor (Latexin, LXN), the co expression of PRR11, CTHRC1 and LXN was detected by immunohistochemistry. The correlation between PRR11 and CTHRC1 and LXN expression was analyzed. Methods 1. tissue samples and patient information were selected from the pathological database to select 216 cases of gastric cancer patients in accordance with the following conditions: from January 2001 to December 2005, surgery was performed in the general surgery of the PLA Changhai Hospital of Shanghai. Patients with gastric cancer treated by resection were followed up to December 2010. HE staining sections and tissue paraffin blocks were selected from the tissue specimen bank. Two experienced experts in science were independently identified as gastric cancer. The clinicopathological parameters of the patients were age, sex, tumor size, T staging, N staging, TNM staging and tumor differentiation. All the samples were obtained, all received the consent of the patient, signed the informed consent, and obtained the Tissue microarray, TMA and Immunohistochemistry (Immunohistochemistry, IHC) of the ethics committee of Changhai Hospital (.2., Immunohistochemistry, IHC), which were performed by two experienced pathological professionals on all the HE stained tissue slides in all patients. Diagnosis and identification, identified as gastric cancer tissue, and marked on a representative site. A tissue column with a diameter of 1.5mm was removed from each of the donor wax blocks, and the.4um thick slices in the receptor paraffin were placed on the slides coated with 3- amino propyl triethoxy silane (3-aminopropyltriethoxysilane, APTS). The antibody was diluted as follows: Anti - PRR11 (1:100 dilution), anti -LXN (1:100 dilution); anti -CTHRC1 (1:100 dilution). Streptomycin peroxidase (Streptavidin-perosidase, S-P) immunohistochemical staining made the antibody binding tissue color, hematoxylin re dyeing.PRR11, LXN, CTHRC1 immunohistochemical staining was observed by two independent experts under Olympus CX31 microscope. The number of PRR11 protein staining (cytosolic Brown) was 10% that was positive.3. cell line and culture conditions found in 5 GC cell lines (SGC7901, MKN45, MKN28, HGC27, MGC803) and found that PRR11 protein expressed.GC cell lines in dur Burke modified Igor medium (Dulbecco modified) plus 10% fetal bovine serum Serum, FBS), and at 37 C, cultured in a incubator containing 5%CO2, the cell coverage rate is about 80%, and the.4. through the SH RNA lentivirus knockdown gastric cancer cells express the short hairpin containing the target PRR11 (Short hairpin RNA, 5 '). Constructed, 6ug/ml polyamines were used to transfect gastric cancer cells. The transfected gastric cancer cells were selected with purinamycin, and Immunoblotting (Western blot) was used to detect the effect of PRR11 knockdown on.5. cell proliferation analysis and cloning form analysis for the density of 5000 cells per pore, and the stable transfected PRR11-sh RNA thin. Cell (PRR11-KO cells) or empty carrier negative control cells (PRR11-NC cells) were inoculated in 96 orifice plates, each set of three vice pores, using cell count box (Cell counting Kit-8, CCK8), to calculate the cell proliferation rate by calculating the absorbance of 24 hours and 48 hours. With the density of 500 cells per pore, PRR11-KO cells and PRR11-NC cells were inoculated with the density of PRR11-KO cells and PRR11-NC cells. In the 6 orifice, each three holes. After two weeks of incubation, methanol / acetone (1:1) was fixed, crystal violet was stained, and the number of more than 50 cell clones,.6. Western blot, was prepared to prepare 5 kinds of gastric cancer cell lines, using Rabbit anti human PRR11, CTHRC1, LNX polyclonal antibody (1:1000 dilute release), and the combination of horseradish peroxidase (Horse radish peroxidase, HRP). The Goat anti rabbit Ig G antibody (1:10000 dilution) was used as two resistance and beta -actin as the internal parameter. According to the manufacturer's instructions, the Amersham enhanced chemiluminescence system was used for protein development and.7. real-time quantitative reverse transcriptase PCR (Q RT-PCR) analysis. The real-time quantitative reverse transcriptase was carried out by SYBR1 Premix Ex. The synthase chain reaction (Quantitative Real-time reverse transcription PCR, Q RT-PCR), glyceraldehyde -3- phosphate dehydrogenase (Glyceraldehyde-3-phosphate dehydrogenase, GAPDH) as the internal parameter.PCR program is 95 centigrade 1 minutes, then 95 centigrade 15 seconds, 60 centigrade 31 seconds, a total of 40 cycles. PCR detection and analysis. Primers are used as follows: PRR11: before 5 '-CGT ATC TGC CAC CGA GAA CTT-3', 5 '-GAG ATG GTC. C7901 cells, PRR11-KO cells, tumor formation and growth ability change.PRR11-KO and PRR11-NC cells (1 x 106 cells 0.1ml PBS) were injected into the right axillary of female Balb/c nude mice in 4 weeks of age respectively. The tumor diameter was measured every three days. All nude mice were killed at 2 weeks after transplantation. The tumor was collected and the tumor volume was measured, and V=0.52 (long x width * high).9. statistics Analysis and statistical analysis used the analysis of the relationship between SPSS 13 software.PRR11 protein expression and the clinicopathological parameters of gastric cancer using the chi 2 test (chi-squared distribution). According to the expression of PRR11 protein, the gastric cancer patients were stratified and the Kaplan-Meier method was used for survival analysis. The single factor and multifactor analysis were based on the Cox proportional risk regression model. The measurement data were presented with mean standard deviation. The measurement data were analyzed by Student 't- test.P? 0.05. The results were statistically significant. 1. the expression of PRR11 protein in gastric carcinoma and normal gastric mucosa was significantly different. The expression of PRR11 protein was related to the clinicopathological parameters of gastric cancer, and 216 cases of gastric cancer were diagnosed as gastric cancer. The results showed that the immune staining of PRR11 protein in normal gastric mucosa was low, and the immune staining in some gastric carcinoma tissues was high. In 216 cases of gastric carcinoma, 107 cases (49.5%) PRR11 protein expression was positive, 109 cases (50.5%) PRR11 protein expression negative.Q RT-PCR and Western blot The analysis further demonstrated that PRR11 m RNA and protein were highly expressed in gastric cancer tissue samples compared with the corresponding normal gastric mucosa. In 5 gastric cancer cell lines, such as SGC7901, MKN45, MKN28, HGC27 and MGC803, we found that PRR11 protein expressed the relationship between the expression of.PRR11 protein expression and the clinicopathological parameters of multiple gastric cancer. The expression of PRR11 protein was not associated with age, sex, tumor size and N staging, but was significantly related to T staging, degree of tumor differentiation, and TNM staging. The results suggest that the high expression of PRR11 protein may be associated with the up regulation of.2.PRR11 expression related to the invasion and metastasis of gastric cancer and the decrease of the survival of 216 patients with gastric cancer, and 122 cases are in the patients with gastric cancer. During the follow-up period, the median survival period (Median survival time, MST) was 74.5 months for gastric cancer patients with negative.PRR11 protein expression for 51.5 months, while the median survival period of the gastric cancer patients with PRR11 positive expression was 46.4 months. In addition, the single variable COX regression analysis showed the size of tumor, the T staging of the tumor, regional lymph node metastasis, TN. M staging, tumor differentiation and PRR11 expression were significantly related to the total survival of patients (Overall survival, OS). Multivariate analysis showed that tumor size, tumor differentiation, and PRR11 expression were independent prognostic factors of gastric cancer patients. The patients were divided into two subgroups of I/II phase and III/IV phase, and the effect of PRR11 expression on the survival time of each subgroup was analyzed. The results showed that in the two subgroups of I/II and III/IV phase, compared with PRR11 non expression patients, the prognosis of PRR11 high expression patients was poor (P0.05).3.PRR11 low expression and proliferation of SGC7901 gastric cancer cells in vitro, the decrease of clone formation and the slow growth of tumor in vivo was associated with the lentivirus PRR11 sh RNA stable silent SGC7901 cell line in the SGC7901 cell line. .Western blot method confirmed that PRR11 low expression SGC7901 cell line (PRR11-KO) was successfully established. The expression of PRR11 in PRR11-KO cells decreased significantly. The low expression of PRR11 was correlated with the proliferation and cloning of SGC7901 cell lines in vitro. Similarly, the downregulation of PRR11 led to the growth of SGC7901 cell lines in nude mice to delay the low expression of.4.PRR11. After the downregulation of CTHRC1 expression and up regulation of LXN expression, we found that the decrease of PRR11 expression led to the downregulation of CTHRC1m RNA expression and the up regulation of LXN m RNA expression. Further, Western blot analysis was used to explore whether these proteins decreased with the expression of CTHRC1 protein and LXN protein antibody. The results showed that the expression of CTHRC1 protein in PRR11-KO cells was down regulated and the expression of LXN protein was up-regulated in the PRR11-NC cells. The expression of CTHRC1 protein in gastric carcinoma tissue was positively correlated with the expression of PRR11 protein (r=0.299, p0.001), LNX protein expression and PRR11 protein table in gastric carcinoma. Negative correlation (r=-0.188, p=0.005). There was no correlation between the expression of CTHRC1 protein and the expression of LXN protein (r=-0.042, p=0.539). Conclusion the high expression of 1.PRR11 protein in gastric cancer tissues is low in normal gastric mucosa, and is related to the pathological parameters of gastric cancer differentiation, T staging, TNM staging and so on. This suggests that the high expression of PRR11 protein may be associated with gastric cancer. The.2. multivariate analysis showed that the size, differentiation and PRR11 protein expression of gastric cancer were the total number of gastric cancer patients.
【學(xué)位授予單位】：第三軍醫(yī)大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2015
【分類號】：R735.2

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