本文選題：肝癌 + NKT淋巴�。� 參考：《青島大學(xué)》2017年碩士論文

【摘要】：目的:通過(guò)研究肝癌組織中自然殺傷T細(xì)胞(Natural killer T cells,NKT)的數(shù)量分布變化與肝癌轉(zhuǎn)移的相關(guān)性,進(jìn)一步明確NKT細(xì)胞與肝癌疾病進(jìn)展期的變化規(guī)律,為肝癌的預(yù)后判斷與治療提供科學(xué)依據(jù),增加NKT細(xì)胞在肝臟疾病中的臨床數(shù)據(jù),為進(jìn)一步探索肝臟微環(huán)境與NKT細(xì)胞之間的相互影響,提供實(shí)驗(yàn)依據(jù)并探索肝癌免疫治療新方法。研究對(duì)象及方法:1、研究對(duì)象:收集青島大學(xué)附屬醫(yī)院肝膽外科、普外科2016.01~2016.05年間手術(shù)切除并經(jīng)病理證實(shí)的肝癌標(biāo)本共64例(術(shù)前未行放、化療),標(biāo)本根據(jù)全國(guó)肝癌協(xié)會(huì)會(huì)議擬定的分期為標(biāo)準(zhǔn)進(jìn)行臨床分期,均經(jīng)4g/L的中性甲醛固定,常規(guī)石蠟包埋,4um厚常規(guī)切片,蘇木精-伊紅(HE)染色和復(fù)核病理診斷。另取非肝癌手術(shù)的正常肝組織15例作為對(duì)照。2、方法:選擇小鼠抗人CD3單抗和小鼠抗人CD56單抗,采用雙重免疫組織化學(xué)SP法對(duì)切片進(jìn)行抗原染色,DAB和AEC顯色劑分別顯色后,光學(xué)顯微鏡下分別計(jì)數(shù)在正常肝臟組織,臨床不同分期肝癌組織中NKT淋巴細(xì)胞的數(shù)量及分布情況,運(yùn)用統(tǒng)計(jì)學(xué)方法分析NKT淋巴細(xì)胞的數(shù)量分布變化與肝癌細(xì)胞遷移的相關(guān)性。切片中能同時(shí)表達(dá)CD3、CD56抗原分子的淋巴細(xì)胞為陽(yáng)性NKT細(xì)胞。結(jié)果:1、15例正常肝臟組織NKT細(xì)胞單個(gè)高倍視野平均數(shù)為18.3571 8.3265,且NKT細(xì)胞多集中在門(mén)管區(qū)微血管和肝血竇內(nèi)。2、28例Ⅰa-Ⅱa期肝癌組織NKT細(xì)胞單個(gè)高倍視野平均數(shù)為9.2438 2.5894,隨病程進(jìn)展肝臟內(nèi)微環(huán)境的變化NKT細(xì)胞數(shù)量逐漸下降。3、36例Ⅱb-Ⅲb肝癌組織NKT細(xì)胞單個(gè)高倍視野平均數(shù)為7.6587 5.9854。晚期肝癌NKT細(xì)胞數(shù)量進(jìn)一步下降,視野內(nèi)NKT數(shù)量很少。4、肝癌組織中(未分期)自然殺傷性T淋巴細(xì)胞的數(shù)量明顯低于正常肝組織內(nèi)NKT細(xì)胞的數(shù)量,具有明顯差異P0.05(P=0.037)5、不同分期肝癌中Ⅰa期-Ⅱa期與Ⅱb期-Ⅲb期的NKT細(xì)胞數(shù)量有明顯差異P0.05(P=0.025),且癌細(xì)胞侵襲肝臟的程度越高,NKT細(xì)胞數(shù)量越少。結(jié)論:(1)利用不同的免疫細(xì)胞化學(xué)顯色劑,分別顯色NKT細(xì)胞膜上表達(dá)的CD3和CD56抗原,對(duì)NKT細(xì)胞進(jìn)行識(shí)別和定位。(2)正常肝臟組織NKT細(xì)胞細(xì)胞多集中在門(mén)管區(qū)微血管內(nèi)和肝血竇內(nèi)。(3)肝癌組織中(未分期)自然殺傷性T淋巴細(xì)胞的數(shù)量明顯低于正常肝組織內(nèi)NKT細(xì)胞的數(shù)量。(4)NKT細(xì)胞數(shù)量隨肝癌病程進(jìn)展逐漸下降。癌細(xì)胞侵襲肝臟的程度越高,NKT細(xì)胞數(shù)量越少。
[Abstract]:Objective: to study the relationship between the quantity distribution of natural killer T cells and the metastasis of hepatocellular carcinoma (HCC), and to study the relationship between NKT cells and the progression of HCC. It can provide scientific basis for prognosis and treatment of liver cancer, increase the clinical data of NKT cells in liver diseases, further explore the interaction between liver microenvironment and NKT cells, provide experimental basis and explore a new method of immunotherapy for liver cancer. Subjects and methods: study subjects: we collected 64 specimens of hepatobiliary surgery from the affiliated Hospital of Qingdao University, which were surgically resected and pathologically proved by general surgery during the period of 2016.01and 2016.05. All the specimens were fixed by neutral formaldehyde of 4g/L, paraffin-embedded 4um thick conventional sections, hematoxylin-eosin (HEH) staining and pathological diagnosis. Another 15 cases of normal liver tissue were taken as control group. Methods: mouse anti-human CD3 monoclonal antibody and mouse anti-human CD56 monoclonal antibody were selected, and the sections were stained by antigen staining and AEC respectively by double immunohistochemistry SP method. The number and distribution of NKT lymphocytes in normal liver tissues and clinical HCC tissues were counted under optical microscope. The correlation between the number distribution of NKT lymphocytes and the migration of HCC cells was analyzed by statistical method. The lymphocytes which could express CD3 + CD56 antigen were positive NKT cells. Results the average of single high-power field of NKT cells in 15 normal liver tissues was 18.3571 8.3265, and the average of NKT cells was 9.2438 2.5894 in 28 patients with stage 鈪，

本文編號(hào)：1913912