本文選題：心肌細胞培養(yǎng)液 + 鼻咽癌細胞�。� 參考：《瀘州醫(yī)學院》2012年碩士論文

【摘要】：本課題組前期研究體外已經(jīng)證實心肌細胞培養(yǎng)液能明顯抑制鼻咽癌細胞的生長，對非腫瘤細胞無抑制作用，推測心肌細胞培養(yǎng)液中可能含有一種或幾種安全有效的抑瘤活性物質(zhì)，該抑瘤活性無濃度依賴性。體內(nèi)實驗研究發(fā)現(xiàn)心肌細胞培養(yǎng)液能抑制S180小鼠移植瘤生長，不影響小鼠的正常生長；在CNE2裸鼠移植瘤模型得到了相同的答案，證實了該抑瘤活性物質(zhì)的安全性和有效性，并進一步研究發(fā)現(xiàn)其抑瘤機制與誘導腫瘤細胞凋亡有關，同腫瘤微血管的生成無關；經(jīng)葡聚糖凝膠層析分離提純及質(zhì)譜鑒定推測心肌細胞培養(yǎng)液中抑瘤活性成分中可能含有EIF-5A或該蛋白的裂解片段。 目的：通過體外抑瘤實驗探討心肌細胞培養(yǎng)液（myocardial cells culturemedium，CMCM）對人鼻咽癌低分化鱗癌細胞系生長的影響，并對其抑瘤機制進行探討。 方法：分別于CMCM作用于CNE-2細胞12小時、24小時、36小時、48小時、60小時、72小時采用MTT法檢測細胞的存活情況，同時以順鉑（DDP1.26μg/ml）作為陽性對照，RPMI1640培養(yǎng)基作為陰性對照，，計算出細胞存活率，繪制生長曲線；并在作用24小時時采用流式細胞儀檢測各組CNE-2細胞的周期分布及凋亡情況。 結果：作用12小時后CMCM組與DDP組細胞存活率（x±s，%）分別為87.27±6.99、89.88±0.72，與陰性對照組100.00±4.06相比便存在顯著差異（P0.05）。隨著時間的延長，差異逐漸增大，72小時后差異達最大，CMCM組與DDP組細胞存活率（x|-±s，%）分別為21.45±2.45、12.72±1.52，與陰性對照組100.00±10.69相比，差異均具有統(tǒng)計學意義。各實驗組分別作用于鼻咽癌細胞24小時后，流式細胞儀檢測細胞周期分布及凋亡情況。CMCM組G0/G1期細胞比率（x|-|-±s，%）為68.70±3.40，與陰性對照組50.16±2.92相比，差異具有統(tǒng)計學意義，DDP組G0/G1期細胞比率為47.97±1.87，與陰性對照組相比，差異無統(tǒng)計學意義。DDP組G2/M期細胞比率（x|-±s，%）為22.10±2.67，明顯高于陰性對照組6.60±2.03（P0.05），CMCM組G2/M期細胞比率為8.10±1.78，與陰性對照組相比，差異無統(tǒng)計學意義。細胞凋亡率（x|-±s，%）CMCM組與DDP組分別為39.80±2.08、5.86±1.66，與陰性對照組0.11±0.05相比，差異均具有統(tǒng)計學意義。 結論：心肌細胞培養(yǎng)液對鼻咽癌細胞CNE-2生長的抑制作用具有時間依賴性。心肌細胞培養(yǎng)液鼻咽癌細胞CNE-2生長的抑制作用機制可能與誘導細胞凋亡及G0/G1期細胞阻滯有關。
[Abstract]:Our previous studies have demonstrated in vitro that cardiomyocyte culture medium can significantly inhibit the growth of nasopharyngeal carcinoma cells, but has no inhibitory effect on non-tumor cells. It is speculated that there may be one or several safe and effective anti-tumor active substances in cardiomyocyte culture medium, which is not concentration-dependent. In vivo experiments showed that cardiomyocyte culture medium could inhibit the growth of transplanted tumor in S180 mice without affecting the normal growth of mice. The same answer was obtained in CNE2 nude mice model of transplanted tumor, which confirmed the safety and efficacy of the anti-tumor active substance. Furthermore, it was found that the mechanism of tumor inhibition was related to the induction of apoptosis of tumor cells, but not to the formation of tumor microvessels. The anti-tumor components of cardiomyocyte culture medium may contain the fragment of EIF-5A or this protein in the cardiomyocyte culture medium by Glucan gel chromatography separation and mass spectrometry. Objective: to investigate the effect of myocardial cells culturemedia (CMCM) on the growth of poorly differentiated human nasopharyngeal carcinoma (NPC) cell lines in vitro and its mechanism. Methods: CNE-2 cells were treated with CMCM for 12 hours, 24 hours, 36 hours, 48 hours, 60 hours and 72 hours, respectively. MTT assay was used to detect the survival rate of CNE-2 cells, and cisplatin DDP 1.26 渭 g / ml was used as negative control to calculate the survival rate of CNE-2 cells. The growth curve was drawn and the cell cycle distribution and apoptosis of CNE-2 cells in each group were detected by flow cytometry at 24 hours. Results: the cell survival rate of CMCM group and DDP group was 87.27 鹵6.9 鹵89.88 鹵0.72, which was significantly higher than that of negative control group (100.00 鹵4.06). With the prolongation of time, the difference gradually increased 72 hours later, the difference reached the maximum after 72 hours. The cell survival rate of CMCM group and DDP group were 21.45 鹵2.45 鹵1.52 鹵1.52 and 100.00 鹵10.69, respectively. The difference was statistically significant compared with that of negative control group (100.00 鹵10.69). After treated with nasopharyngeal carcinoma cells for 24 hours, the cell cycle distribution and apoptosis were detected by flow cytometry. In CMCM group, the cell ratio of G _ 0 / G _ 1 phase was 68.70 鹵3.40, which was higher than that of negative control group (50.16 鹵2.92). The ratio of G _ 2 / M phase cells in the DDP group was 22.10 鹵2.67, which was significantly higher than that in the negative control group (6.60 鹵2.03P0.05CMCM, 8.10 鹵1.78), compared with the negative control group, the ratio of G _ 2 / M phase cells in the DDP group was significantly higher than that in the negative control group (47.97 鹵1.87), and the ratio of G _ 2 / M phase cells in the DDP group was 22.10 鹵2.67, which was significantly higher than that in the negative control group (8.10 鹵1.78). The difference was not statistically significant. The rate of apoptosis in CMCM group and DDP group was 39.80 鹵2.080.86 鹵1.66, respectively, which was significantly higher than that in negative control group (0.11 鹵0.05). Conclusion: the inhibitory effect of cardiomyocyte culture medium on the growth of nasopharyngeal carcinoma cell line CNE-2 is time dependent. The inhibitory mechanism of CNE-2 growth in cardiomyocyte culture medium may be related to the induction of apoptosis and the arrest of G _ 0 / G _ 1 phase cells.
【學位授予單位】：瀘州醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R739.63

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