亚洲人和高加索人大肠癌细胞体外恶性行为的比较

发布时间：2018-06-19 09:54

本文选题：大肠癌 + CACO2　；参考：《昆明医科大学》2017年硕士论文

【摘要】：[目的]对于NDRG1在亚洲人与高加索人大肠癌中的作用近年来众说纷纭。本文旨在比较亚洲人与高加索人NDRG1基因有无差异,以及分析NDRG1在亚洲人与高加索人大肠癌细胞株体外侵袭中的作用。[方法]利用细胞计数法测定细胞生长速度;通过荧光定量PCR法在mRNA水平比较NDRG1在亚洲人和高加索人大肠癌细胞中的表达;24-Transwell法检测亚洲人和高加索人大肠癌细胞侵袭和迁移能力;用流式细胞术检测亚洲人和高加索人大肠癌细胞的大小、细胞周期、DNA含量及NDRG1蛋白含量等。[结果]分析CACO2与SNU-C1细胞生长曲线得出CACO2细胞的增殖速度比SNU-C1细胞快,且差异有统计学意义(P0.05);利用PCR法检测CACO2与SNU-C1两株细胞,通过比对核酸序列得出两株细胞所扩增出来的序列均与NDRG1 一致;利用QPCR法检测CACO2与SNU-C1中mRNA含量得出 CACO2 与 SNU-C1 的 NDRG1 的 mRNA 水平比值为 0.4061,SNU-C1 的 NDRG1的mRNA水平为CACO2的2.46倍;Transwell法检测细胞的侵袭与迁移能力,发现CACO2细胞侵袭能力在24h、48h和72h各时间点均高于SNU-C1,但24h两组细胞之间差异无统计学意义(P=0.083),而48h和72h两个时间点两组细胞之间差异有统计学意义(P=0.006,P=0.000),而迁移实验中CACO2组除24h组迁移能力比SNU-C1组低之外,其它时间点迁移能力均高于SNU-C1组,且两组细胞之间差异有统计学意义(P=0.038,P=0.045,P=0.012);利用流式细胞术检测两株细胞大小,经FSC分析发现CACO2平均体积明显大于SNU-C1,从SSC分析得到SNU-C1平均颗粒度大于CACO2、SNU-C1细胞表面的皱折度、细胞内亚细胞器、颗粒的数目等均大于CACO2;利用流式细胞术检测细胞周期,结果显示两种细胞各间期之间差异无统计学意义(G1间期P=0.546,S间期P=0.119,G2间期P=0.071);利用流式细胞术检测两株细胞之间NDRG1蛋白含量,结果显示CACO2细胞MEAN值为36.3,SNU-C1细胞MEAN值为86.5;用流式细胞术检测两株细胞NDRG1的DNA含量,结果显示MEAN值:CACO2细胞为45,SNU-C1细胞为50.2。[结论]1.亚洲人和高加索人NDRG1相同,但在大肠癌中表达水平不同。2.同等条件下高加索人大肠癌细胞株CACO2的体外侵袭和迁移能力强于亚洲人大肠癌细胞株SNU-C1的。
[Abstract]:[objective] the role of NDR G 1 in colorectal cancer in Asians and Caucasians has been controversial in recent years. The purpose of this study was to compare the difference of NDRG1 gene between Asians and Caucasians and to analyze the role of NDRG1 in invasion of colorectal cancer cell lines in vitro. [methods] Cell growth rate was measured by cell counting method. The expression of NDRG1 in colorectal cancer cells of Asian and Caucasian human was compared by fluorescence quantitative PCR method and the ability of invasion and migration of colorectal cancer cells in Asia and Caucasus was detected by 24-Transwell method. The cell size, cell cycle DNA content and NDRG1 protein content were measured by flow cytometry in Asian and Caucasian colorectal cancer cells. [results] by analyzing the growth curve of CACO _ 2 and SNU-C1 cells, we found that the proliferation rate of CACO _ 2 cells was faster than that of SNU-C1 cells, and the difference was statistically significant (P _ (0.05), CACO _ 2 and SNU-C1 cell lines were detected by PCR. The nucleic acid sequence of the two cells was identical to that of NDRG1. The mRNA level ratio of NDRG1 of CACO _ 2 to SNU-C1 was 0.4061 and the mRNA level of NDRG1 of SNU-C1 was 2.46 times higher than that of CACO _ 2. It was found that the invasive ability of CACO _ 2 cells was higher than that of SNU-C1 at 24 h and 72 h, but there was no significant difference between the two groups at 24 h and 72 h, but there was significant difference between the two groups at 48 h and 72 h. Except that the migration ability of 24h group was lower than that of SNU-C1 group, The migration ability of SNU-C1 group was higher than that of SNU-C1 group at other time points, and the difference between the two groups was statistically significant. FSC analysis showed that the mean volume of CACO _ 2 was significantly larger than that of SNU-C1.The average particle size of SNU-C1 was larger than that of CACO _ 2C _ 2 SNU-C1 cells, and the suborganelles and the number of particles were all larger than that of CACO _ 2. Flow cytometry was used to detect cell cycle. The results showed that there was no significant difference between the two cell interphase. The protein content of NDRG1 was detected by flow cytometry. The results showed that the Mean value of CACO _ 2 cells was 36.3 SNU-C1 was 86.5, and the DNA content of NDRG1 was detected by flow cytometry. The results showed that the value of Mei _ 2: CACO _ 2 cells was 45 SNU-C1 cells, and the DNA content of NDRG1 cells was 50.2. [conclusion] 1. Asian and Caucasian NDRG1 are the same, but the expression level of NDRG1 is different in colorectal cancer. The invasion and migration ability of caucasian colorectal cancer cell line CACO _ 2 was stronger than that of Asian colorectal cancer cell line SNU-C1 under the same conditions.
【学位授予单位】：昆明医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.34

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