本文選題：大腸癌 + CACO2��； 參考：《昆明醫(yī)科大學》2017年碩士論文

【摘要】：[目的]對于NDRG1在亞洲人與高加索人大腸癌中的作用近年來眾說紛紜。本文旨在比較亞洲人與高加索人NDRG1基因有無差異,以及分析NDRG1在亞洲人與高加索人大腸癌細胞株體外侵襲中的作用。[方法]利用細胞計數法測定細胞生長速度;通過熒光定量PCR法在mRNA水平比較NDRG1在亞洲人和高加索人大腸癌細胞中的表達;24-Transwell法檢測亞洲人和高加索人大腸癌細胞侵襲和遷移能力;用流式細胞術檢測亞洲人和高加索人大腸癌細胞的大小、細胞周期、DNA含量及NDRG1蛋白含量等。[結果]分析CACO2與SNU-C1細胞生長曲線得出CACO2細胞的增殖速度比SNU-C1細胞快,且差異有統(tǒng)計學意義(P0.05);利用PCR法檢測CACO2與SNU-C1兩株細胞,通過比對核酸序列得出兩株細胞所擴增出來的序列均與NDRG1 一致;利用QPCR法檢測CACO2與SNU-C1中mRNA含量得出 CACO2 與 SNU-C1 的 NDRG1 的 mRNA 水平比值為 0.4061,SNU-C1 的 NDRG1的mRNA水平為CACO2的2.46倍;Transwell法檢測細胞的侵襲與遷移能力,發(fā)現CACO2細胞侵襲能力在24h、48h和72h各時間點均高于SNU-C1,但24h兩組細胞之間差異無統(tǒng)計學意義(P=0.083),而48h和72h兩個時間點兩組細胞之間差異有統(tǒng)計學意義(P=0.006,P=0.000),而遷移實驗中CACO2組除24h組遷移能力比SNU-C1組低之外,其它時間點遷移能力均高于SNU-C1組,且兩組細胞之間差異有統(tǒng)計學意義(P=0.038,P=0.045,P=0.012);利用流式細胞術檢測兩株細胞大小,經FSC分析發(fā)現CACO2平均體積明顯大于SNU-C1,從SSC分析得到SNU-C1平均顆粒度大于CACO2、SNU-C1細胞表面的皺折度、細胞內亞細胞器、顆粒的數目等均大于CACO2;利用流式細胞術檢測細胞周期,結果顯示兩種細胞各間期之間差異無統(tǒng)計學意義(G1間期P=0.546,S間期P=0.119,G2間期P=0.071);利用流式細胞術檢測兩株細胞之間NDRG1蛋白含量,結果顯示CACO2細胞MEAN值為36.3,SNU-C1細胞MEAN值為86.5;用流式細胞術檢測兩株細胞NDRG1的DNA含量,結果顯示MEAN值:CACO2細胞為45,SNU-C1細胞為50.2。[結論]1.亞洲人和高加索人NDRG1相同,但在大腸癌中表達水平不同。2.同等條件下高加索人大腸癌細胞株CACO2的體外侵襲和遷移能力強于亞洲人大腸癌細胞株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.
【學位授予單位】：昆明醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R735.34

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本文編號：2039485