| 李 超,袁 鹏,颜昭勇,张建省,张洪新,牟 佼.节律基因CRY1在肝癌中的表达及对肝癌细胞生长的作用[J].现代生物医学进展英文版,2017,17(33):6432-6436. |
| 节律基因CRY1在肝癌中的表达及对肝癌细胞生长的作用 |
| Expression of CRY1 in Hepatocellular Carcinoma and its Effects on the Growth of Hepatocellular Carcinoma Cells |
| Received:July 22, 2017 Revised:August 18, 2017 |
| DOI：10.13241/j.cnki.pmb.2017.33.007 |
| 中文关键词: CRY1 生物节律 增殖 凋亡 肝细胞肝癌 |
| 英文关键词: CRY1 circadian rhythm Proliferation Apoptosis HCC |
| 基金项目:国家自然科学基金项目（81572304，81600478） |
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| 中文摘要: |
| 摘要 目的：探讨节律基因CRY1（cryptochrome circadian clock 1）在肝癌组织中的表达情况及对肝癌细胞生长的作用。方法：利用实时荧光定量PCR、Western blotting及免疫组织化学染色法，检测CRY1在肝癌组织及细胞中的表达；利用肝癌公共数据信息，分析和验证CRY1在肝癌组织中的表达；真核表达质粒将肝癌细胞CRY1过表达后，采用MTS和Annexin V-FITC/PI 双染法，分别检测CRY1对细胞增殖和凋亡的影响。结果：CRY1在癌组织中阳性率为57.5%（23/40），癌旁组织中阳性率为87.5%（35/40），癌中CRY1表达低于癌旁的占80%（32/40），与癌旁组织比较，CRY1蛋白在肝癌组织中的表达明显降低，差异有统计学意义（t=7.53，P<0.001）。CRY1在3个肝癌公共数据信息显示，与对应癌旁组织相比，其在癌组织中表达水平显著降低，差异均具有统计学意义（TCGA：413.62 vs 335.17，P=0.0155；GSE22058：397.67 vs 360.08，P=0.0207；GSE25097：1.627 vs 1.433，P<0.0001）。同时CRY1在被检测的4株肝癌细胞系中表达也显著低于永生化的正常肝细胞。与对照组比，过表达CRY1组细胞增殖能力明显降低，两组间差异有统计学意义（1.100 vs 0.662，P<0.0001）；与对照组比，过表达CRY1组细胞凋亡显著增加（凋亡细胞百分比：7.9 vs 17.5，P=0.0014）。结论：节律基因CRY1在肝癌中低表达，同时CRY1抑制肝癌细胞增殖和促进细胞凋亡。 |
| 英文摘要: |
| ABSTRACT Objective: To explore the expression of CRY1 in hepatocellular carcinoma (HCC) and its effects on the growth of HCC cells. Methods: The expression of CRY1 in HCC was detected by real-time quantitative PCR (qRT-PCR), Western blotting and immuno- histochemical (IHC) staining. Using public dataset from HCC patients to analyze and validate the expression of CRY1. The effects of CRY1 on cell proliferation and apoptosis were detected by MTS and Annexin V-FITC/PI double staining after enhanced the expression of CRY1 in HCC cells by using eukaryotic expression plasmid. Results: The positive rate of CRY1 expression was 57.5% (23/40) in tu- mor tissues and was 87.5% (35/40) in peritumor tissues. The expression of CRY1 in tumor tissues was lower than that in peritumor tis- sues to 80%(32/40). Compared with peritumor tissues, the expression of CRY1 protein in HCC tissues was significantly decreased, the difference was statistically significant(t=7.53, P<0.001). Compared with the corresponding peritumor tissues, the expression of CRY1 showed a significant reduction in tumor tissues from three public dataset of HCC patients, and the differences were statistically signifi- cant (TCGA: 413.62 vs 335.17, P=0.0155; GSE22058: 397.67 vs 360.08, P=0.0207; GSE25097: 1.627 vs 1.433, P<0.0001). Meanwhile, the expression of CRY1 in four HCC cell lines was significantly lower than that in normal hepatocytes. Compared with the control group, the proliferation ability of the CRY1 overexpression group was significantly decreased, and the difference between the two groups was statistically significant (1.100 vs 0.662, P<0.0001). Compared with the control group, apoptosis in the CRY1 overexpression group was significantly increased (percentage of apoptotic rate: 7.9 vs 17.5, P=0.0014). Conclusion: The expression of CRY1 is low in HCC, and CRY1 can inhibit proliferation and promote apoptosis of HCC cells. |
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