本文選題：空間環(huán)境 + B16細(xì)胞��； 參考：《昆明醫(yī)學(xué)院》2006年碩士論文

【摘要】：研究目的： 1．通過動(dòng)物實(shí)驗(yàn)觀察空間誘變小鼠黑色素瘤B16細(xì)胞株的成瘤性，探索通過體內(nèi)實(shí)驗(yàn)篩選空間誘變B16細(xì)胞的方法，尋找免疫原性發(fā)生變異的陽性細(xì)胞株。 2．觀察可疑陽性空間誘變小鼠黑色素瘤B16細(xì)胞細(xì)胞骨架和細(xì)胞表面結(jié)構(gòu)，觀察可疑變異細(xì)胞株的細(xì)胞骨架和表面結(jié)構(gòu)與對(duì)照細(xì)胞株的差異。 3．檢測(cè)體內(nèi)篩選獲得的實(shí)驗(yàn)陽性細(xì)胞株的體外培養(yǎng)生長(zhǎng)曲線和細(xì)胞周期分布等生物學(xué)特性，觀察可疑變異細(xì)胞株的生物學(xué)特性與對(duì)照細(xì)胞株的差異。 4．觀察可疑變異細(xì)胞株基因表達(dá)的改變，尋找與黑色素瘤發(fā)生及轉(zhuǎn)移密切相關(guān)的基因。 研究方法： 1．將B16細(xì)胞株應(yīng)用細(xì)胞低溫長(zhǎng)期生存系統(tǒng)，搭載于我國(guó)第20顆返回式衛(wèi)星，返地后進(jìn)行單克隆化，從得到的110株單克隆太空誘變B16細(xì)胞株中隨機(jī)選取4株，常規(guī)培養(yǎng)傳代5代后和對(duì)照細(xì)胞株同時(shí)分別接種C57BL/6J小鼠，腹腔接種組觀察生存期，皮下接種組14天后取血、處死，記錄瘤重，脾臟重，胸腺重，并進(jìn)行血清細(xì)胞因子檢測(cè)和瘤體的病理學(xué)檢測(cè)。 2．應(yīng)用激光共聚焦顯微鏡觀察篩選所得陽性細(xì)胞株與對(duì)照細(xì)胞株分別用熒光素標(biāo)記的鬼筆環(huán)肽染色，觀察細(xì)胞骨架比較異同。 3．應(yīng)用原子力顯微鏡(AFM)對(duì)篩選所得陽性細(xì)胞株和對(duì)照細(xì)胞株分別成像，分別觀察空間誘變細(xì)胞株和對(duì)照組細(xì)胞所得圖像的差異。 4．MTT法檢測(cè)篩選所得陽性細(xì)胞株細(xì)胞生長(zhǎng)曲線與對(duì)照組進(jìn)行比較。 5．應(yīng)用流式細(xì)胞儀(FCM)檢測(cè)篩選所得陽性細(xì)胞株和對(duì)照細(xì)胞株的細(xì)胞周期分布。
[Abstract]:Objectives of the study: 1. The tumorigenesis of murine melanoma B16 cell line induced by space was observed through animal experiments. The method of screening space mutated B16 cell line in vivo was explored to find out the positive cell line with variant immunogenicity. 2. The cytoskeleton and surface structure of murine melanoma B16 cells induced by suspected positive space mutagenesis were observed, and the difference of cytoskeleton and surface structure between murine melanoma B16 cell line and control cell line was observed. 3. The biological characteristics of culture growth curve and cell cycle distribution in vitro were detected, and the differences between the cell line and the control cell line were observed. 4. To observe the changes of gene expression in suspected variant cell lines and to search for genes closely related to melanoma development and metastasis. Research methods: 1. The B16 cell line was carried on the 20th recoverable satellite in China with a cryogenic long-term survival system. After returning to earth, the B16 cell line was monoclonal transformed. Four of the 110 Monoclonal Space mutated B16 cell lines were randomly selected. C57BL/6J mice were inoculated with conventional culture for 5 passages and control cell lines at the same time. The survival time was observed in the intraperitoneal inoculation group. The blood was taken from the subcutaneous inoculation group after 14 days. The tumor weight, spleen weight, thymus weight were recorded, and the tumor weight, spleen weight and thymus weight were recorded. Serum cytokines and tumor pathology were detected. 2. Laser confocal microscopy was used to observe the difference of cytoskeleton between the positive cell line and the control cell line. 3. Atomic force microscopy (AFM) was used to image the positive cell lines and control cell lines, respectively, and to observe the difference between the space mutagenesis cell lines and the control cells. 4.MTT assay was used to detect the cell growth curve of positive cell line and to compare with the control group. 5. Flow cytometry (FCM) was used to detect the cell cycle distribution of positive cell lines and control cell lines.
【學(xué)位授予單位】：昆明醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2006
【分類號(hào)】：R85

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 李金國(guó),姜國(guó)勇,王培生,張健,蔣興村;谷子種子經(jīng)高空氣球搭載后的遺傳變異研究[J];航天醫(yī)學(xué)與醫(yī)學(xué)工程;1999年05期

2 李金國(guó),劉敏,王培生,張健,薛淮,郭亞華;空間條件對(duì)番茄誘變作用及遺傳的影響[J];航天醫(yī)學(xué)與醫(yī)學(xué)工程;2000年02期

3 李源祥,李金國(guó),楊新喜,蔣興村,華育堅(jiān),黎二妹;贛早秈47號(hào)親本空間誘變的性狀變異研究[J];航天醫(yī)學(xué)與醫(yī)學(xué)工程;2002年02期

4 印紅,謝申義,章光明,謝申猛;空間飛行對(duì)紅曲霉菌產(chǎn)量的影響[J];航天醫(yī)學(xué)與醫(yī)學(xué)工程;2003年05期

5 祁章年，陳湄，李向高;近地軌道衛(wèi)星艙內(nèi)輻射測(cè)量[J];航天醫(yī)學(xué)與醫(yī)學(xué)工程;1994年02期

6 賈士芳，許怡，郭興華，劉志恒，阮繼生;空間條件對(duì)芽孢桿菌的超氧化物歧化酶和其它性質(zhì)的影響[J];航天醫(yī)學(xué)與醫(yī)學(xué)工程;1995年01期

7 李金國(guó)，蔣興村，，段留生;空間條件對(duì)棘孢小單孢菌的誘變效應(yīng)[J];航天醫(yī)學(xué)與醫(yī)學(xué)工程;1995年02期

8 李向高，陳湄，祁章年，王衛(wèi)紅，房興業(yè)，賈向紅;衛(wèi)星艙內(nèi)電離輻射強(qiáng)度的動(dòng)態(tài)變化[J];航天醫(yī)學(xué)與醫(yī)學(xué)工程;1996年01期

9 賈士芳,郭興華,周志宏,劉志恒,李雪東,楊靜華;利用返回式科學(xué)衛(wèi)星選育優(yōu)良雙歧桿菌[J];航天醫(yī)學(xué)與醫(yī)學(xué)工程;1996年06期

10 李金國(guó),王培生,韓東,陳芳遠(yuǎn),鄧力平,郭亞華;高空氣球飛行對(duì)水稻、青椒的誘變效應(yīng)(英文)[J];航天醫(yī)學(xué)與醫(yī)學(xué)工程;1997年02期

本文編號(hào)：1920422