下咽癌FADU細胞系中CD133陽性細胞的基礎研究
發(fā)布時間：2018-10-22 07:07
【摘要】： [研究背景與目的] 下咽癌是一種頭頸部惡性腫瘤,是上消化道及上呼吸道惡性程度最高的腫瘤之一,其發(fā)生的機制并不清楚。預后差的主要原因包括局部復發(fā)、易發(fā)生淋巴結(jié)轉(zhuǎn)移及遠處轉(zhuǎn)移。由于臨床后果差,對患者生命及生活質(zhì)量影響嚴重,是當前耳鼻咽喉-頭頸外科的研究重點。 腫瘤干細胞是近年來對腫瘤發(fā)生發(fā)展過程的一種新的理論觀點,新觀點認為腫瘤組織中只有極少量的腫瘤細胞有干細胞樣作用,它們是一群數(shù)量較少但具有無限自我更新潛能并能促使腫瘤形成的細胞,在腫瘤生長中起著決定性作用。這類細胞有較強的抗拒放療及化療的作用,并可能與免疫逃逸,生長信號異常傳遞等多種腫瘤的重要生物學特點有關。目前已從腦腫瘤,白血病,乳腺癌,喉癌,肺癌等多種腫瘤組織中提取到了腫瘤干細胞,并進行了相關細胞學及分子生物學研究。從當前的研究結(jié)果來看,不同的腫瘤中,盡量其腫瘤干細胞生物學特性有共同點,如無限自我更新潛能,分化能力等,但腫瘤干細胞其表面標志物是不盡相同的,這提示我們需要對不同的腫瘤進行相應研究,了解不同腫瘤中腫瘤干細胞的標志物特點。 近年來,許多研究證實,CD133是腫瘤干細胞的重要標志物之一。CD133是一種分子量為120kDa,具有5個跨膜區(qū)的糖蛋白,屬于一種早期抗原。在2000年英國舉行的第7屆國際人類白細胞分化抗原研討會上被正式命名為CD133。它是一種正常的造血干細胞表面標志物,它的一個顯著特點就是：CD133的表達隨著細胞的分化迅速下調(diào),選擇性地在骨髓和外周血造血干細胞及內(nèi)皮祖細胞表達,但在成熟內(nèi)皮細胞上不表達,是一個重要的分離和鑒定造血干或祖細胞的分子標志。 侵襲能力是惡性腫瘤的標志性行為特點之一,是當前研究惡性腫瘤行為特點的一個重要方向。目前利用Transwell小室是體外研究腫瘤細胞侵襲和轉(zhuǎn)移行為的簡便且快速的方法。 本研究擬以CD133為分選標志物,從人類FADU細胞系中分選出CD133陽性細胞,研究其數(shù)量分布特點,形態(tài)學特點,并擬通過應用Transwell小室的研究方法,研究在FADU細胞系中CD133陽性細胞的侵襲能力,從而為未來完善對CD133陽性細胞擁有的細胞學特點研究提供相應的細胞學實驗室基礎。 第一部分 下咽癌FADU細胞系中CD133表達情況研究 [目的]研究下咽癌FADU細胞系中CD133表達情況。 [方法]對下咽癌FADU細胞進行培養(yǎng)和傳代,分別應用免疫細胞化學和流式細胞儀的方法對下咽癌FADU細胞系中CD133表達情況進行研究。 [結(jié)果]免疫細胞化學的方法發(fā)現(xiàn),在下咽癌FADU細胞中CD133陽性細胞僅占少數(shù),而在利用流式細胞儀的方法研究后發(fā)現(xiàn),FADU細胞中CD133陽性細胞比例僅為2.01%。 [結(jié)論]CD133陽性細胞數(shù)量在FADU細胞系中占少數(shù)。 第二部分 下咽癌FADU細胞系中CD133陽性細胞的分選 [目的]應用可靠的分選手段,從下咽癌FADU細胞系中分選CD133陽性細胞并鑒定。 [方法]對下咽癌FADU細胞進行培養(yǎng)和傳代,應用免疫磁珠分選技術進行CD133陽性細胞的分選。 [結(jié)果]應用免疫磁珠分選技術后,可很好地分選出CD133陽性細胞,其分選后CD133陽性細胞純度可達91.2%。 [結(jié)論]免疫磁珠分選技術可很好地應用于下咽癌FADU細胞系中CD133陽性細胞的分選。 第三部分 下咽癌FADU細胞系中CD133陽性細胞的侵襲能力研究 [目的]研究下咽癌FADU細胞系中CD133陽性細胞的侵襲能力。 [方法]對應用免疫磁珠分選技術進行分選后的CD133陽性細胞、陰性細胞分別應用Transwell小室進行侵襲能力研究,倒置相差顯微鏡下照相,計數(shù)遷移到上室基質(zhì)膜外表面的細胞數(shù)目,以能穿過基質(zhì)膜的細胞數(shù)目多少表示細胞的侵襲能力強弱。每個樣本計數(shù)5個視野(×200)的細胞數(shù),取其平均值進行比較。 [結(jié)果]與CD133陰性細胞相比,CD133陽性細胞透膜數(shù)量較陰性細胞明顯高,其穿過基質(zhì)膜的細胞數(shù)分別為54.0±2.3,17.0±1.1,其差異具有統(tǒng)計學意義,P0.01。 [結(jié)論]在下咽癌FADU細胞系中CD133陽性細胞比陰性細胞侵襲能力強。
[Abstract]:[Background and Purpose of Study] Hypopharyngeal carcinoma is a head and neck malignant tumor, which is one of the most malignant tumors of upper gastrointestinal tract and upper respiratory tract. It is not clear that the main causes of poor prognosis include local recurrence, easy lymph node metastasis, distant metastasis. Due to poor clinical consequences, the patient's life and quality of life seriously affect the patient's life and quality of life. It is the current Otolaryngology-Head and Neck Surgery. Tumor stem cells are a new theoretical point in the development of tumorigenesis in recent years. The new viewpoint holds that only a very small number of tumor cells in tumor tissue have stem cell-like function, they are a group of small numbers but have unlimited self-renewal potential and can A cell that promotes the formation of a tumor in a tumor. It plays a decisive role in long term. These cells have strong resistance to radiation therapy and chemotherapy, and may be associated with various tumors such as immune escape, abnormal transmission of growth signals, and so on. The tumor stem cells are extracted from various tumor tissues such as brain tumor, leukemia, breast cancer, laryngeal cancer, lung cancer and the like, From the current research results, the biological characteristics of tumor stem cells in different tumors have commonalities, such as infinite self-renewal potential, differentiation ability, etc., but the surface of tumor stem cells Markers are different, which suggests that we need to study different tumors to learn about tumors in different tumors The marker features of stem cells. In recent years, many studies have demonstrated that CD133 It is one of the important markers of tumor stem cells. CD133 is a 120kDa molecular weight with 5 transmembrane domains. Glycoprotein of the region belonging to an early antigen. The 7th International Human Leukocyte Differentiation Antigen, held in England in 2000 CD133, a normal hematopoietic stem cell surface marker, is characterized by a rapid down-regulation of CD133 expression with the differentiation of cells, selectively in bone marrow and peripheral blood hematopoietic stem cells and endothelial progenitor cells Da, but not expressed on mature endothelial cells, is an important separation and Identification of molecular markers of hematopoietic stem or progenitor cells. The invasive ability is one of the characteristics of malignant tumors. At present, it is an important direction to study the characteristics of malignant tumor behavior. In vitro, Transwell chamber is used to study swelling in vitro. In this study, CD133 positive cells were selected from human FADU cell lines by CD133 as a sorting marker, and the number and morphology of CD133 positive cells were studied in human FADU cell lines. To investigate the invasion ability of CD133 positive cells in FADU cell lines, and to improve CD133 positive cells in the future. intracellular The cytological feature study provides a corresponding cytological laboratory basis. CD1 in the first hypopharyngeal carcinoma FADU cell line[Objective] To study the expression of CD133 in FADU cell line of hypopharyngeal carcinoma. The expression of CD133 in the FADU cell line of hypopharyngeal carcinoma was studied by immunocytochemistry and flow cytometry. The cells accounted for only a small number of cells, whereas after study using a flow cytometer, FADU was found Cells in C The proportion of D133 positive cells was only 2. 01%.[Conclusion] The number of CD133 positive cells is a few in FADU cell lines. Sorting of CD133-positive cells in hypopharyngeal carcinoma FADU cell line[Objective] To apply a reliable sorting device from hypopharyngeal carcinoma In FADU cell line, CD133 positive cells were sorted and identified.[Methods] FADU cells of hypopharyngeal carcinoma were cultured and passaged, and immunomagnetic beads were applied. Separation of CD133 positive cells by sorting technique.[Results] After using immunomagnetic beads sorting technique, it can be very good. to be selected from the ground CD133 positive cells, the purity of CD133 positive cells after sorting can reach 1. 2%.[Conclusion] Immunomagnetic bead sorting technique can be well applied to hypopharynx Isolation of CD133-positive cells in carcinoma FADU cell lines. Study on invasion ability of CD133-positive cells in the third hypopharyngeal carcinoma FADU cell line[Objective] To study CD13 in the hypopharyngeal carcinoma FADU cell line 3. The invasive ability of positive cells.[Methods] Applied immune magnetism the CD133 positive cells and the negative cells after sorting by the bead sorting technique are respectively used as Tr. An analysis of invasive ability in an answell cell, the number of cells transferred to the outer surface of the upper chamber substrate film by the inversion phase contrast microscope, and the number of cells that can pass through the substrate film indicates the cell's invasion ability. Each sample counts 5 fields of view (Table 2 00) Number of cells to be compared.[Results] Negative with CD133
【學位授予單位】：山東大學
【學位級別】：博士
【學位授予年份】：2010
【分類號】：R739.63
本文編號：2286455
[Abstract]:[Background and Purpose of Study] Hypopharyngeal carcinoma is a head and neck malignant tumor, which is one of the most malignant tumors of upper gastrointestinal tract and upper respiratory tract. It is not clear that the main causes of poor prognosis include local recurrence, easy lymph node metastasis, distant metastasis. Due to poor clinical consequences, the patient's life and quality of life seriously affect the patient's life and quality of life. It is the current Otolaryngology-Head and Neck Surgery. Tumor stem cells are a new theoretical point in the development of tumorigenesis in recent years. The new viewpoint holds that only a very small number of tumor cells in tumor tissue have stem cell-like function, they are a group of small numbers but have unlimited self-renewal potential and can A cell that promotes the formation of a tumor in a tumor. It plays a decisive role in long term. These cells have strong resistance to radiation therapy and chemotherapy, and may be associated with various tumors such as immune escape, abnormal transmission of growth signals, and so on. The tumor stem cells are extracted from various tumor tissues such as brain tumor, leukemia, breast cancer, laryngeal cancer, lung cancer and the like, From the current research results, the biological characteristics of tumor stem cells in different tumors have commonalities, such as infinite self-renewal potential, differentiation ability, etc., but the surface of tumor stem cells Markers are different, which suggests that we need to study different tumors to learn about tumors in different tumors The marker features of stem cells. In recent years, many studies have demonstrated that CD133 It is one of the important markers of tumor stem cells. CD133 is a 120kDa molecular weight with 5 transmembrane domains. Glycoprotein of the region belonging to an early antigen. The 7th International Human Leukocyte Differentiation Antigen, held in England in 2000 CD133, a normal hematopoietic stem cell surface marker, is characterized by a rapid down-regulation of CD133 expression with the differentiation of cells, selectively in bone marrow and peripheral blood hematopoietic stem cells and endothelial progenitor cells Da, but not expressed on mature endothelial cells, is an important separation and Identification of molecular markers of hematopoietic stem or progenitor cells. The invasive ability is one of the characteristics of malignant tumors. At present, it is an important direction to study the characteristics of malignant tumor behavior. In vitro, Transwell chamber is used to study swelling in vitro. In this study, CD133 positive cells were selected from human FADU cell lines by CD133 as a sorting marker, and the number and morphology of CD133 positive cells were studied in human FADU cell lines. To investigate the invasion ability of CD133 positive cells in FADU cell lines, and to improve CD133 positive cells in the future. intracellular The cytological feature study provides a corresponding cytological laboratory basis. CD1 in the first hypopharyngeal carcinoma FADU cell line[Objective] To study the expression of CD133 in FADU cell line of hypopharyngeal carcinoma. The expression of CD133 in the FADU cell line of hypopharyngeal carcinoma was studied by immunocytochemistry and flow cytometry. The cells accounted for only a small number of cells, whereas after study using a flow cytometer, FADU was found Cells in C The proportion of D133 positive cells was only 2. 01%.[Conclusion] The number of CD133 positive cells is a few in FADU cell lines. Sorting of CD133-positive cells in hypopharyngeal carcinoma FADU cell line[Objective] To apply a reliable sorting device from hypopharyngeal carcinoma In FADU cell line, CD133 positive cells were sorted and identified.[Methods] FADU cells of hypopharyngeal carcinoma were cultured and passaged, and immunomagnetic beads were applied. Separation of CD133 positive cells by sorting technique.[Results] After using immunomagnetic beads sorting technique, it can be very good. to be selected from the ground CD133 positive cells, the purity of CD133 positive cells after sorting can reach 1. 2%.[Conclusion] Immunomagnetic bead sorting technique can be well applied to hypopharynx Isolation of CD133-positive cells in carcinoma FADU cell lines. Study on invasion ability of CD133-positive cells in the third hypopharyngeal carcinoma FADU cell line[Objective] To study CD13 in the hypopharyngeal carcinoma FADU cell line 3. The invasive ability of positive cells.[Methods] Applied immune magnetism the CD133 positive cells and the negative cells after sorting by the bead sorting technique are respectively used as Tr. An analysis of invasive ability in an answell cell, the number of cells transferred to the outer surface of the upper chamber substrate film by the inversion phase contrast microscope, and the number of cells that can pass through the substrate film indicates the cell's invasion ability. Each sample counts 5 fields of view (Table 2 00) Number of cells to be compared.[Results] Negative with CD133
【學位授予單位】：山東大學
【學位級別】：博士
【學位授予年份】：2010
【分類號】：R739.63
【參考文獻】
相關期刊論文 前2條
1 王春梅,趙育楨,高凌寒,劉芳莉,李鈺,李璞;應用重組基底膜侵襲技術對人肺腺癌細胞侵襲能力的研究[J];哈爾濱醫(yī)科大學學報;2000年05期
2 ;Inhibitory effects of antisense RNA of HAb18G/CD147 on invasion of hepatocellular carcinoma cells in vitro[J];World Journal of Gastroenterology;2003年10期
，本文編號：2286455
本文鏈接：http://sikaile.net/yixuelunwen/yank/2286455.html
最近更新
教材專著
