本文選題：JAK2V617F + 髓增殖性腫瘤　； 參考：《承德醫(yī)學(xué)院》2016年碩士論文

【摘要】：目的:觀察促血管新生因子:血管內(nèi)皮生長(zhǎng)因子(VEGF)、缺氧誘導(dǎo)因子1α(HIF-1α)及血管新生因子抑制因子:與張力蛋白同源的10號(hào)染色體缺失的磷酸酶基因(PTEN)在Janus蛋白酪氨酸激酶2(JAK2)點(diǎn)突變(JAK2V617F)陽(yáng)性骨髓增殖性腫瘤(MPN)患者骨髓中的表達(dá)水平,探討它們與血管新生之間的相互關(guān)系。方法:1本研究所用的52例石蠟標(biāo)本均取自保定市第一醫(yī)院病理科,選取自2012年1月至2014年10月保定市第一醫(yī)院門診及住院收治的42例JAK2V617F表達(dá)陽(yáng)性的MPN患者及10例特發(fā)性免疫性血小板減少性紫癜(ITP)患者。MPN患者包括真性紅細(xì)胞增多癥(PV)10例,原發(fā)性血小板增多癥(ET)17例,原發(fā)性骨髓纖維化(PMF)15例,其中男18例,女24例,年齡32~75歲,中位年齡57歲。ITP患者包括男5例,女5例,中位年齡40歲。所選取患者的診斷均符合《血液病診斷及療效標(biāo)準(zhǔn)》,患者均知情同意及經(jīng)保定市第一醫(yī)院倫理委員會(huì)批準(zhǔn)。2實(shí)驗(yàn)分組:JAK2V617F陽(yáng)性的MPN患者中初治組(初診未治療組)25例;治療組(該組患者均應(yīng)用干擾素-α2b(IFN-α2b)肌肉或皮下注射,300萬(wàn)U/次,3~7次/周,至少應(yīng)用6個(gè)月以上,聯(lián)合或不聯(lián)合應(yīng)用羥基脲調(diào)整外周血白細(xì)胞、血紅蛋白及血小板數(shù)量。)17例。10例特發(fā)性免疫性血小板減少性紫癜(ITP)患者作為對(duì)照組。3通過實(shí)時(shí)熒光定量PCR檢測(cè)MPN患者JAK2V617F突變量(突變型與野生型JAK2比值)。4應(yīng)用免疫組化方法檢測(cè)初治組、治療組及對(duì)照組患者骨髓病理切片p-JAK2、VEGF、HIF-1α、PTEN的蛋白表達(dá)水平及CD105標(biāo)記的微血管密度(MVD)。5所有數(shù)據(jù)均采用SPSS 17.0統(tǒng)計(jì)分析軟件,計(jì)量資料以(x±s)表示,兩組均數(shù)間的比較采用t檢驗(yàn),多組均數(shù)比較采用方差分析,兩兩比較采用q檢驗(yàn)。spearman等級(jí)相關(guān)分析各變量之間的相關(guān)性。jak2v617f突變陽(yáng)性中jak2v617f突變量、vegf、hif-1α、mvd表達(dá)的比較及四指標(biāo)與臨床特征(血栓栓塞發(fā)生率)的關(guān)系分析采用χ2檢驗(yàn);p0.05為差異有統(tǒng)計(jì)學(xué)意義。結(jié)果:1mpn患者的jak2v617f突變量pcr結(jié)果顯示:42例mpn患者在初次診治時(shí)均經(jīng)實(shí)時(shí)熒光定量pcr檢測(cè)顯示jak2v617f突變?yōu)殛?yáng)性,經(jīng)過治療后jak2v617f突變轉(zhuǎn)陰性者2例,其余40例mpn患者jak2v617f突變量在26.8%-75.4%之間。其中初治組25例,jak2v617f突變量為46.17%±19.32%,干擾素治療組17例,jak2v617f突變量為22.69%±12.64%,前者明顯高于后者(p0.05)。2免疫組織化學(xué)染色檢測(cè)p-jak2、vegf、hif-1α、pten、mvd在mpn患者中的表達(dá)水平p-jak2、vegf、hif-1α及pten蛋白均表達(dá)于細(xì)胞質(zhì),其中p-jak2、vegf、hif-1α三者骨髓細(xì)胞中陽(yáng)性率在初治組(82.41%±11.65%;64.72%±25.01%;45.12%±20.28%)中表達(dá)最高,其次為治療組(60.93%±20.57%;36.58%±15.95%;32.15%±14.27%),對(duì)照組(43.05%±12.59%;25.69%±13.75%;25.07%±15.49%)最低,差異均有統(tǒng)計(jì)學(xué)顯著性(p0.01);mvd在初治組(26.58%±5.93%)中顯著高于治療組(15.86%±4.27%)及對(duì)照組(10.76%±4.01%),pten蛋白在初治組(24.78%±14.86%)中表達(dá)最低,其次為治療組(38.72%±18.25%),對(duì)照組表達(dá)(55.05%±21.17%)最高,差異均有統(tǒng)計(jì)學(xué)意義(p0.01)。3jak2v617f突變量與p-jak2、vegf、hif-1α、pten、mvd相關(guān)分析spearman等級(jí)相關(guān)分析顯示mpn患者的jak2v617f突變量和p-jak2、vegf、mvd正相關(guān)(r=0.739,p0.01、r=0.589,p0.05、r=0.577,p0.05),與hif-1α無(wú)明顯相關(guān)(p0.05),與pten負(fù)相關(guān)(r=-0.508,p0.05)。pten蛋白表達(dá)與mvd負(fù)相關(guān)(r=-0.584,p0.05)。以jak2v617f/jak2比值50%為界值分為50%與≥50%兩組,其中50%組26例,≥50%組16例。結(jié)果顯示jak2v617f/jak250%的患者其p-jak2、vegf、hif-1α及mvd(65.15%±17.59%、43.26%±18.67%、33.27%±15.26%、19.16%±5.34%)的水平均明顯低于jak2v617f/jak2≥50%組(89.76%±20.56%、69.87%±27.32%、49.51%±22.65%、34.19%±6.57%),pten在jak2v617f/jak2比值50%患者的水平(34.42%±19.76%)要明顯高于jak2v617f/jak2比值≥50%(22.68%±13.12%)患者(p均0.05)。初治組中(46.17%±19.32%;82.41%±11.65%;64.72%±25.01%;45.12%±20.28%;26.58%±5.93%)jak2v617f突變量及p-jak2、vegf、hif-1α、mvd的水平均明顯高于ifn-α2b治療組(22.69%±12.64%;60.93%±20.57%;64.72%±25.01%;32.15%±14.27%;15.86%±4.27%;15.86%±4.27%)及對(duì)照組(0;43.05%±12.59%;25.69%±13.75%;25.07%±15.49%;10.76%±4.01%)(p均0.05);而pten與上述相反,在初治組(24.78%±14.86%)中表達(dá)最低,其次為治療組(15.86%±4.27%),在對(duì)照組(10.76%±4.01%)中表達(dá)最高(p均0.05)。4jak2v617f及mvd與血栓栓塞的相關(guān)性42例患者,初治組25例,血栓栓塞15例,血栓發(fā)生率為60.00%;治療組17例,再發(fā)血栓栓塞4例(既往發(fā)生血栓者不計(jì)在內(nèi)),血栓發(fā)生率為23.53%;后者明顯低于前者(χ2=5.43,p0.05)。42例mpn患者中19例患者發(fā)生血栓,血栓發(fā)生率為45.24%,其中26例jak2v617f/jak2比值50%組,發(fā)生血栓栓塞者8例,血栓發(fā)生率為30.77%,16例≥50%組,發(fā)生血栓栓塞者11例,血栓發(fā)生率為68.75%,后者明顯高于前者(χ2=5.77,p0.05)。42例患者總vegf為53.40%±21.97%,其中vegf53.40%患者24例,血栓栓塞7例,血栓發(fā)生率為33.33%,vegf53.40%患者18例,血栓栓塞12例,血栓發(fā)生率為66.67%,后者明顯高于前者(χ2=5.84,p0.05)。42例患者總hif-1α為39.46%±18.08%,其中hif-1α39.46%患者22例,血栓栓塞6例,血栓發(fā)生率為31.82%,hif-1α39.46%患者20例,血栓栓塞13例,血栓發(fā)生率為60.00%,后者明顯高于前者(χ2=6.02,p0.05)。42例患者總mvd為22.24±7.49,其中mvd22.24患者23例,血栓栓塞7例,血栓發(fā)生率為為30.43%,mvd22.24患者19例,血栓栓塞12例,血栓發(fā)生率為63.16%,后者明顯高于前者(χ2=4.50,P0.05)。結(jié)果表明,經(jīng)干擾素治療后,隨著JAK2V617F突變量的減低,患者血栓栓塞發(fā)生率亦減低;JAK2V617F突變量愈高,MVD愈高,VEGF表達(dá)越高,HIF-1α表達(dá)越高,患者血栓栓塞發(fā)生率愈高,呈正相關(guān)關(guān)系。結(jié)論:1 PTEN、VEGF、HIF-1α與JAK2V617F共同參與了骨髓增殖性腫瘤患者血管新生。2 JAK2V617F陽(yáng)性MPN患者在應(yīng)用IFN-α2b治療后JAK2V617F突變量明顯減少,部分患者轉(zhuǎn)陰性,IFN-α2b能夠抑制MPN患者JAK2V617F突變量。3 VEGF、HIF-1α及MVD在JAK2V617F陽(yáng)性患者中表達(dá)增加,而PTEN表達(dá)減低。4 JAK2V617F基因突變量越高者VEGF、HIF-1α、MVD越高則血管新生越明顯,而與PTEN表達(dá)負(fù)相關(guān)。5 JAK2V617F突變量愈高,VEGF表達(dá)越高,HIF-1α表達(dá)越高,骨髓微血管密度愈高,患者發(fā)生血栓風(fēng)險(xiǎn)則愈高,隨著JAK2V617F突變量的減低,患者血栓栓塞發(fā)生率亦減低。
[Abstract]:Objective: To observe vascular endothelial growth factor: vascular endothelial growth factor (VEGF), hypoxia inducible factor 1 alpha (HIF-1 alpha) and angiogenesis factor inhibitory factor: phosphatase gene (PTEN), which is homologous to tension protein 10 chromosome deletion (PTEN) in Janus protein tyrosine kinase 2 (JAK2) point mutation (JAK2V617F) positive marrow proliferative tumor (MPN) patients' bone marrow The relationship between them and angiogenesis was discussed. Methods: 1 paraffin specimens from 1 studies were taken from the pathology department of the first hospital in Baoding. From January 2012 to October 2014, 42 cases of MPN patients with positive MPN and 10 idiopathic immune blood were treated in the outpatient and hospitalized Hospital of the first hospital of the city. .MPN patients included 10 cases of real erythrocytosis (PV), 17 cases of primary thrombocythemia (ET) and 15 cases of primary myelofibrosis (PMF), of which 18 cases were male, 24 women, age 32~75 years, and 57 year old.ITP patients, including 5 men, 5 women, and middle age 40. The diagnosis of the selected patients was in line with < blood. The diagnosis and efficacy criteria of liquid disease, the patients' informed consent and the.2 experimental group approved by the ethics committee of the first hospital of Baoding: 25 patients with JAK2V617F positive MPN patients (first diagnosed untreated group); the treatment group (all the patients were treated with interferon - alpha 2b (IFN- alpha 2b) muscle or subcutaneous injection, 3 million U/ times, 3~7 times per week, at least 6 months. Above, combined or non combined use of hydroxyurea in peripheral blood leukocytes, hemoglobin and platelet counts.) 17 cases of.10 patients with idiopathic thrombocytopenic purpura (ITP) as control group,.3 by real-time fluorescence quantitative PCR detection of MPN patients JAK2V617F process variable (mutate and wild type JAK2 ratio).4 application immunization method The p-JAK2, VEGF, HIF-1, PTEN protein expression level and the microvascular density (MVD).5 of CD105 markers in the treatment group and the control group were measured with SPSS 17 statistical analysis software, and the measurement data were (x + s). The comparison between the two groups was carried out by t test, and the multiple groups were compared with variance analysis. 22 compared the correlation analysis of the correlation.Jak2v617f mutation between the variables of.Spearman and the correlation analysis between the variables, the comparison of the VEGF, HIF-1 alpha, MVD expression and the relationship between the four indexes and the clinical characteristics (thromboembolism incidence) by the Q test. The analysis of the relationship between the four indexes and the clinical characteristics (thromboembolism incidence) was analyzed by the chi 2 test; P0.05 was statistically significant. The result: JAK2V617F of the 1mpn patients. The results of sudden variable PCR showed that 42 cases of MPN patients were detected by real-time fluorescence quantitative PCR at the first time of diagnosis and JAK2V617F mutation was positive. After treatment, 2 cases were JAK2V617F mutation negative, and the other 40 cases of MPN patients were between 26.8%-75.4%. The initial treatment group was 25 cases, JAK2V617F process variable was 46.17% + 19.32%, interferon treatment. In group 17, the JAK2V617F process variable was 22.69% + 12.64%. The former was significantly higher than that of the latter (P0.05).2 immunohistochemical staining for p-jak2, VEGF, HIF-1 alpha, PTEN, MVD in MPN patients, p-jak2, VEGF, HIF-1 alpha and protein were expressed in the cytoplasm, and the positive rate in bone marrow cells was 82.41% + 11. (82.41% + 11.). The expression of 65%, 64.72% + 25.01%, 45.12% + 20.28%) was the highest, followed by the treatment group (60.93% + 20.57%; 36.58% + 15.95%; 32.15% + 14.27%), and the control group (43.05% + 12.59%; 25.69% + 20.28%) was the lowest, and the difference was statistically significant (P0.01); MVD was significantly higher in the primary treatment group than the treatment group and the control group. .76% + 4.01%), the expression of PTEN protein was the lowest in the primary treatment group (24.78% + 14.86%), followed by the treatment group (38.72% + 18.25%) and the control group (55.05% + 21.17%), the difference was statistically significant (P0.01).3jak2v617f abrupt variable and p-jak2, VEGF, HIF-1 a, PTEN, MVD correlation analysis of Spearman grade correlation analysis showed JAK2V617F process variables of MPN patients The positive correlation with p-jak2, VEGF, MVD (r=0.739, P0.01, r=0.589, P0.05, r=0.577, P0.05) was not significantly correlated with HIF-1 alpha (P0.05), and was negatively correlated with PTEN negative correlation (r=0.739). The value of the ratio 50% was divided into 50% and more than 50% groups, including 50% groups 26 cases, and 50% groups 16 cases. The levels of p-jak2, VEGF, HIF-1 alpha and MVD (65.15% + 17.59%, 43.26% + 18.67%, 33.27% + 15.26%, 19.16% + 5.34%) were significantly lower than those of jak2v617f/jak2 > 50% group (89.76% + 20.56%, 69.87% + 27.32%, 43.26% + 33.27%). The level of PTEN at JAK2V617F /jak2 ratio was significantly higher than that of jak2v617f/. The JAK2 ratio was more than 50% (22.68% + 13.12%) (P 0.05). In the primary treatment group (46.17% + 19.32%; 82.41% + 11.65%; 64.72% + 25.01%; 45.12% + 20.28%; 26.58% + 5.93%), the level of p-jak2, VEGF, HIF-1 alpha and MVD was significantly higher than that of ifn- alpha 2b; 86% + 4.27%) and the control group (0; 43.05% + 12.59%; 25.69% + 13.75%; 25.07% + 15.49%; 10.76% + 4.01%) (P 0.05); while PTEN was the lowest in the primary treatment group (24.78% + 14.86%), followed by the treatment group, the highest (P mean).4jak2v617f and MVD and thromboembolism were expressed in the control group. In the patients, there were 25 cases in the primary treatment group, 15 cases of thromboembolism and 60% thrombus, 17 cases in the treatment group, 4 cases of recurrent thromboembolism (not included in the previous thrombus), the thrombus incidence was 23.53%, the latter was significantly lower than the former (x 2=5.43, P0.05).42 cases MPN patients with thrombosis, the incidence of thrombosis was 45.24%, of which 26 cases jak2v617f/jak2 ratio. In the 50% group, thromboembolism occurred in 8 cases, thrombus incidence was 30.77%, 16 cases were more than 50%, thromboembolism occurred in 11 cases, thrombus incidence was 68.75%, the latter was significantly higher than the former (x 2=5.77, P0.05).42 patients with total VEGF 53.40% + 21.97%, 24 cases of vegf53.40%, thromboembolism 7 cases, thrombus incidence of 33.33%, 18 cases of vegf53.40% patients, 18 cases, vegf53.40% patients, Thromboembolism in 12 cases, the incidence of thrombus was 66.67%, the latter was significantly higher than the former (x 2=5.84, P0.05).42 cases, the total HIF-1 alpha was 39.46% + 18.08%, including 22 cases of HIF-1 alpha 39.46%, thromboembolism in 6 cases, thrombus incidence of 31.82%, 20 cases of HIF-1 a 39.46%, thromboembolism in 13 cases, thrombus incidence of 60%, the latter was significantly higher than the former (x 2=6.02, P) 0.05) the total MVD of.42 patients was 22.24 + 7.49, of which 23 cases of mvd22.24, 7 thromboembolism, 30.43% thrombus, 19 cases of mvd22.24, 12 thromboembolism and 63.16% thrombus, the latter was significantly higher than that of the former (x 2=4.50, P0.05). The results showed that after interferon treatment, thrombus thrombus was decreased along with the decrease of JAK2V617F process variable. Thrombus thrombus The higher the incidence of plug is, the higher the JAK2V617F process, the higher the MVD, the higher the expression of VEGF, the higher the expression of HIF-1 a, the higher the incidence of thromboembolism in the patients. Conclusion: 1 PTEN, VEGF, HIF-1 alpha and JAK2V617F are involved in the new.2 JAK2V617F positive MPN patients in the patients with proliferative tumor of the bone marrow. IFN- alpha 2b could inhibit.3 VEGF of JAK2V617F process in MPN patients, HIF-1 A and MVD increased in JAK2V617F positive patients, while PTEN expression reduced the greater of.4 genes, the higher the higher then the higher then the more obvious, and the negative correlation with the expression of 2B. The higher the quantity, the higher the expression of VEGF, the higher the expression of HIF-1 a, the higher the density of the bone marrow microvessel, the higher the risk of thrombosis in the patients. The incidence of thromboembolism is also decreased with the decrease of the variable of JAK2V617F.
【學(xué)位授予單位】：承德醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：R733.3

【相似文獻(xiàn)】

相關(guān)會(huì)議論文 前2條

1 陳晶;吳水軍;劉明群;楊楊;張磊;;基于自由加量方式的突變量阻抗判據(jù)測(cè)試研究[A];2012年云南電力技術(shù)論壇論文集（文摘部分）[C];2012年

2 陳晶;吳水軍;劉明群;楊楊;張磊;;基于自由加量方式的突變量阻抗判據(jù)測(cè)試研究[A];2012年云南電力技術(shù)論壇論文集[C];2012年

相關(guān)碩士學(xué)位論文 前3條

1 付建珠;VEGF、HIF-1α、PTEN與JAK2V617F陽(yáng)性骨髓增殖性腫瘤血管新生相關(guān)性研究[D];承德醫(yī)學(xué)院;2016年

2 徐�；�;基于突變量的大型機(jī)組的升壓變壓器主保護(hù)新原理的研究[D];合肥工業(yè)大學(xué);2007年

3 劉大鵬;采樣值突變量距離繼電器的研究與仿真[D];華北電力大學(xué)（北京）;2010年

，

本文編號(hào)：2067415