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目的研究口腔癌过表达蛋白1(oral cancer overexpressed protein 1,ORAOV1)在人结肠癌组织中的表达变化并探讨其对结肠癌细胞增殖能力、细胞周期及早期凋亡的影响。方法收集人结肠癌及癌旁组织标本,分别以qRT-PCR及Western blot检测ORAOV1在mRNA及蛋白水平的表达变化情况;采用RNAi技术获得ORAOV1低表达的Lo Vo及SW480细胞株,并使用Western blot对干扰结果进行鉴定;采用CCK-8检测干扰ORAOV1表达后两种结肠癌细胞增殖能力的变化;并采用流式细胞术检测干扰ORAOV1表达对结肠癌细胞周期及早期凋亡的影响。结果 qRT-PCR及Western blot结果分别显示结肠癌组织中的ORAOV1 mRNA及蛋白水平高于癌旁组织(P<0.05);Western blot检测证实Lo Vo及SW480细胞中ORAOV1干扰成功;在这两种结肠癌细胞株中,CCK-8检测结果显示,干扰ORAOV1可抑制细胞增殖能力;流式细胞术结果显示,干扰ORAOV1可使处于S期比例增加(P<0.05)、早期凋亡比例增加(P<0.05)。结论结肠癌组织中的ORAOV1表达增加,干扰ORAOV1可降低结肠癌细胞的增殖能力,可能与S期阻滞及细胞早期凋亡增加有关。
Objective To investigate the expression changes of oral cancer overexpressed protein 1 (ORAOV1) in human colon cancer and to investigate its effect on the proliferation, cell cycle and early apoptosis of colon cancer cells. Methods Colorectal cancer and para-cancerous tissue specimens were collected and the mRNA and protein levels of ORAOV1 were detected by qRT-PCR and Western blot respectively. The LoOV and SW480 cell lines with low expression of ORAOV1 were obtained by RNAi technique and Western blot The effect of ORAOV1 on the proliferation of two colon cancer cells was detected by CCK-8 assay. The effect of ORAOV1 on the cell cycle and early apoptosis of colon cancer cells was detected by flow cytometry. Results The results of qRT-PCR and Western blot showed that the mRNA and protein levels of ORAOV1 in colorectal cancer tissues were significantly higher than those in para-cancerous tissues (P <0.05). Western blot analysis showed that the ORAOV1 interference in Lo Vo and SW480 cells was successful. The result of flow cytometry showed that the interference with ORAOV1 could increase the proportion in S phase (P <0.05) and increase the proportion of early apoptosis (P <0.05), and CCK-8 results showed that the interference with ORAOV1 could inhibit cell proliferation. 0.05). Conclusion ORAOV1 expression in colon cancer tissues is increased, interference with ORAOV1 can reduce the proliferation of colon cancer cells, which may be related to the arrest of S phase and the increase of early apoptosis of cells.