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纸质出版:2016
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胡梦竹,王娟,张巧,高社干,张功员.永生化食管上皮恶变细胞中核干细胞因子、细胞周期蛋白B1和PTEN的表达[J].陆军军医大学学报(原第三军医大学学报),2016,38(13):1512-1516.
Hu Mengzhu,Wang Juan,Zhang Qiao,Gao Shegan,Zhang Gongyuan.Expression of nucleostemin, Cyclin B1 and PTEN in immortalized esophageal epithelial malignant cells[J].J Amry Med Univ (J Third Mil Med Univ),2016,38(13):1512-1516.
胡梦竹,王娟,张巧,高社干,张功员.永生化食管上皮恶变细胞中核干细胞因子、细胞周期蛋白B1和PTEN的表达[J].陆军军医大学学报(原第三军医大学学报),2016,38(13):1512-1516. DOI:
Hu Mengzhu,Wang Juan,Zhang Qiao,Gao Shegan,Zhang Gongyuan.Expression of nucleostemin, Cyclin B1 and PTEN in immortalized esophageal epithelial malignant cells[J].J Amry Med Univ (J Third Mil Med Univ),2016,38(13):1512-1516. DOI:
目的 观察核干细胞因子(nucleostemin,NS)、细胞周期蛋白B1(Cyclin B1)及抑癌基因PTEN mRNA和蛋白的表达情况。 方法 正常食管上皮细胞(shantou human embryonic esophageal epithelial cell line,SHEE)和永生化食管上皮恶变细胞(shantou human embryonic esophageal carcinoma epithelial cell line,SHEEC)连续传代培养,取对数生长期SHEE和SHEEC细胞,采用RT-PCR及Western blot方法检测细胞中NS、Cyclin B1及PTEN mRNA和蛋白的表达并做相关性分析,MTT法检测抑制NS表达后的细胞增殖活性。 结果 以SHEE细胞中NS、Cyclin B1和PTEN mRNA的表达量为1,SHEEC细胞中对应的相对表达量分别为(2.047±0.507)、(2.214±0.213)及(0.668±0.188),两组比较差异均有统计学意义(P0.05);SHEE及SHEEC细胞中NS、Cyclin B1及PTEN蛋白表达量分别为(1.142±0.113),(3.161±0.108)、(1.585±0.056),(2.173±0.058)及(1.894±0.341),(1.162±0.524),两组比较差异均有统计学意义(P0.05);NS mRNA和蛋白与Cyclin B1 mRNA和蛋白的表达呈正相关(r=0.832,P0.01;r=0.991,P0.05),与PTEN mRNA和蛋白的表达呈负相关(r=-0.924,P0.01;r=-0.769,P0.05);MTT检测结果显示,NS RNA干扰(RNA interference
RNAi)组与空白对照组和阴性对照组在干扰后12 h细胞增殖能力差异无统计学意义(P0.05),而在干扰后24、36、48 h差异有统计学意义(P0.05)。 结论 SHEEC细胞中NS、Cyclin B1 mRNA和蛋白的表达量均高于SHEE,而PTEN的表达量低于SHEE,NS可能对细胞恶变有影响。
Objective To determine the expression of nucleostemin (NS)
Cyclin B1 and tumor suppressor gene
phosphatase and tensin homolog (PTEN) at mRNA and proteins levels in Shantou human embryonic esophageal epithelial cell line (SHEE) and Shantou human embryonic esophageal carcinoma epithelial cell line (SHEEC). Methods The SHEE and SHEEC cells were continuously subcultured
and the cells at the logarithmic growth phase were selected for the expression of NS
Cyclin B1 and PTEN at mRNA and proteins levels by RT-PCR and Western blotting. The levels were analyzed for correlation. Cell viability was detected by MTT assay after interference expression of NS. Results The mRNA expression of NS and Cyclin B1 were 2.047±0.507 and 2.214±0.213 times higher
while that of PTEN was 0.668±0.188 times lower in the SHEEC cells than the SHEE cells (P0.05). Significance differences were also seen in the protein levels of NS
Cyclin B1 and PTEN between the SHEE and SHEEC cells (1.142±0.113 vs 3.161±0.108
1.585±0.056 vs 2.173±0.058
1.894±0.341 vs 1.162±0.524
P0.05). The mRNA and protein expression levels of NS were positively correlated with those of Cyclin B1 (r=0.832
P0.01;r=0.991
P0.01)
and negatively correlated with PTEN (r=-0.924
P0.01;r=-0.769
P0.05). MTT assay showed that there were no significant difference in cell proliferation among NS RNAi group
MOCK group
and NC group at 12 h after RNA interference
but significant differences were seen in 24
36 and 48 h thereafter (P0.05). Conclusion The expression of NS and Cyclin B1 at mRNA and protein levels are higher
while those of PTEN is lower in SHEEC cells than SHEE cells. NS may affect the malignant progression of cells.
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