1.400038 重庆,第三军医大学西南医院全军肝胆外科研究所
2.401120 重庆,重庆医科大学附属第三医院消化疾病中心
Received:06 December 2016,
Revised:2017-03-01,
Published:30 June 2017
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Peng Guo, Shuguo Zheng. Role of long noncoding RNA CCAT2 in modulating biological behaviors of liver cancer cells and the underlying mechanism[J]. Journal of Third Military Medical University, 2017, 39(12): 1213-1218.
Peng Guo, Shuguo Zheng. Role of long noncoding RNA CCAT2 in modulating biological behaviors of liver cancer cells and the underlying mechanism[J]. Journal of Third Military Medical University, 2017, 39(12): 1213-1218. DOI: 10.16016/j.1000-5404.201612030.
目的
2
探讨长链非编码RNA CCAT2(LncRNA CCAT2) 在肝癌中的作用及机制。
方法
2
选取我院2013年1-6月60例肝癌及其癌旁组织,采用实时荧光定量逆转录聚合酶链反应(qRT-PCR)检测其组织中CCAT2的表达,同时培养正常肝细胞L02和人源性肝癌细胞系HepG、H2P、SMMC和HLE细胞,采用qRT-PCR检测细胞中CCAT2的表达,采用siRNA干扰细胞CCAT2表达后,在24、48、72、96 h用CCK-8检测SMMC和HLE细胞增殖;Transwell实验分析细胞迁移和侵袭情况;采用流式细胞仪分析CCAT2对SMMC和HLE细胞周期分布和凋亡的影响;Western blot检测P53、Bcl-2、Caspase-8蛋白的表达。
结果
2
在肝癌组织和肝癌细胞中,CCAT2的表达显著升高(
P
均
<
0.05),CCAT2高表达的患者预后和生存率均较低(
P
均
<
0.05)。肝癌细胞中低表达的CCAT2能显著抑制肝癌细胞增殖、迁移和侵袭(
P
均
<
0.05)。肝癌细胞中低表达的CCAT2也能使肝癌细胞周期停留在G
0
/G
1
期并促进细胞凋亡(
P
均
<
0.05)。采用siRNA干扰细胞CCAT2表达后,肝癌细胞中P53和Caspase-8蛋白的表达显著升高(
P
均
<
0.05),Bcl-2蛋白的表达显著降低(
P
<
0.05)。
结论
2
CCAT2在肝癌组织和细胞系中高表达,siRNA干扰CCAT2表达后能够抑制细胞增殖、迁移、侵袭以及促进肝癌细胞的凋亡,其机制可能通过升高P53和Caspase-8蛋白的表达和降低Bcl-2蛋白的表达有关。
Objective
2
To investigate the role of long noncoding RNA CCAT2 (LncRNA CCAT2) in modulating the proliferation
migration
invasion and apoptosis of liver cancer cells
in vitro
and explore the underlying mechanism.
Methods
2
Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of LncRNA CCAT2 in 60 paired liver cancer and adjacent tissue samples collected from Southwest Hospital between January and June 2013 and in normal human liver cell line L02 and liver cancer cell lines HepG
H2P
SMMC and HLE. SMMC and HLE cells transfected with siRNA-CCAT2 were examined for cell proliferation rate at 24
48
72 and 96hafter transfection using CCK-8 assay; the cell migration and invasion were detected with Transwell chamber test
and the cell cycle distribution and apoptosis were analyzed with flow cytometry. Western blotting was used to detect the expression levels of P53
Bcl-2 and caspase-8 in the transfected SMMC and HLE cells.
Results
2
The expression level of CCAT2 was significantly increased in the liver cancer tissues and cell lines (all
P
<
0.05). The patients withahigh expression level of CCAT2 hadapoorer prognosis andalower overall survival rate (all
P
<
0.05). In SMMC and HLE cells
CCAT2 knockdown by small interfering RNA resulted in cell cycle arrested in G
0
/G
1
phase (
P
<
0.05)
promoted the cell apoptosis (
P
<
0.05)
and enhanced the expression levels of P53 and Caspase-8 but decreased Bcl-2 expression (
P
<
0.05).
Conclusion
2
LncRNA CCAT2 is over-expressed in liver cancer tissue and cell lines. siRNA-mediated CCAT2 knockdown suppresses the proliferation
invasion and migration and promotes apoptosis of liver cancer cells
in vitro
possibly by increasing the expression of P53 and caspase-8 and decreasing the expression of Bcl-2 protein.
Negoi I, Runcanu A, Paun S, et al . Resection of large metachronous liver metastasis with gastric origin: case report and review of the literature[J ] . Cureus, 2016, 8(10): e814. DOI:10.7759/cureus.814.
Bellon E, Gebauer F, Tachezy M, et al . Pancreatic cancer and liver metastases: state of the art[J ] . Updates Surg, 2016, 68(3): 247-251. DOI: 10.1007/s13304-016-0407-7.
Zhou N, Si Z, Li T, et al . Long non-coding RNA CCAT2 functions as an oncogene in hepatocellular carcinoma, regulating cellular proliferation, migration and apoptosis[J ] . Oncol Lett, 2016, 12(1): 132-138. DOI: 10.3892/ol.2016.4580.
Chen X, Liu L, Zhu W. Up-regulation of long non-coding RNA CCAT2 correlates with tumor metastasis and poor prognosis in cervical squamous cell cancer patients[J]. Int J Clin Exp Pathol, 2015, 8(10): 13261-13266.
Redis R S, Vela L E, Lu W, et al . Allele-specific reprogramming of cancer metabolism by the long non-coding RNA CCAT2[J ] . Mol Cell, 2016, 61(4): 520-534. DOI: 10.1016/j.molcel.2016.01.015.
Dickson I. Hepatocellular carcinoma: A role for lncRNA in liver cancer[J]. Nat Rev Gastroenterol Hepatol, 2016, 13(3): 122-123. DOI: 10.1038/nrgastro.2016.21.
Li J, Zhuang C, Liu Y, et al . shRNA targeting long non-coding RNA CCAT2 controlled by tetracycline-inducible system inhibits progression of bladder cancer cells[J ] . Oncotarget, 2016. DOI: 10.18632/oncotarget.8259.
Kasagi Y, Oki E, Ando K, et al . The expression of CCAT2, a novel long noncoding RNA transcript, and rs6983267 single-nucleotide polymorphism genotypes in colorectal cancers[J ] . Oncology, 2017, 92(1): 48-54. DOI: 10.1159/000452143.
Chen S, Wu H, Lv N, et al . LncRNA CCAT2 predicts poor prognosis and regulates growth and metastasis in small cell lung cancer[J ] . Biomedecine & pharmacotherapie, 2016, 82: 583-588. DOI: 10.1016/j.biopha.2016.05.017.
Huang S, Qing C, Huang Z, et al . The long non-coding RNA CCAT2 is up-regulated in ovarian cancer and associated with poor prognosis[J ] . Diagn Pathol, 2016, 11(1): 49. DOI: 10.1186/s13000-016-0499-x.
Weiskirchen R, Tacke F. Immune surveillance of liver cancer in non-alcoholic fatty liver disease: excess lipids cause CD4 T-cells loss and promote hepatocellular carcinoma development[J]. Hepatobiliary Surg Nutr, 2016, 5(5): 433-437. DOI: 10.21037/hbsn.2016.09.10.
Li Y C, Zhao L, Wu J P, et al . Cytokine-induced killer cell infusion combined with conventional treatments produced better prognosis for hepatocellular carcinoma patients with barcelona clinic liver cancer B or earlier stage: A systematic review and meta-analysis[J ] . Cytotherapy, 2016, 18(12): 1525-1531. DOI: 10.1016/j.jcyt.2016.09.002.
Soldera J. Diagnostic and therapeutic approaches to hepatocellular carcinoma: understanding the barcelona Clinic liver cancer protocol[J]. Clin Med Insights Gastroenterol, 2016, 9: 67-71. DOI: 10.4137/cgast.s30190.
Ding L J, Li Y, Wang S D, et al . Long noncoding RNA lncCAMTA1 promotes proliferation and cancer stem cell-like properties of liver cancer by inhibiting CAMTA1[J ] . Int J Mol Sci, 2016, 17(10): E1617. DOI: 10.3390/ijms17101617.
Li S P, Xu H X, Yu Y, et al . LncRNA HULC enhances epithelial-mesenchymal transition to promote tumorigenesis and metastasis of hepatocellular carcinoma via the miR-200a-3p/ZEB1 signaling pathway[J ] . Oncotarget, 2016, 7(27): 42431-42446. DOI: 10.18632/oncotarget.9883.
Wang X, Sun W, Shen W, et al . Long non-coding RNA DILC regulates liver cancer stem cells via IL-6/STAT3 axis[J ] . J Hepatol, 2016, 64(6): 1283-1294. DOI: 10.1016/j.jhep.2016.01.019..
Yu J, Han J, Zhang J, et al . The long noncoding RNAs PVT1 and uc002mbe.2 in sera provide a new supplementary method for hepatocellular carcinoma diagnosis[J ] . Medicine (Baltimore), 2016, 95(31): e4436. DOI: 10.1097/MD.0000000000004436.
Zhu X T, Yuan J H, Zhu T T, et al . Long noncoding RNA glypican 3 (GPC3) antisense transcript 1 promotes hepatocellular carcinoma progression via epigenetically activating GPC3[J ] . FEBS J, 2016, 283(20): 3739-3754. DOI: 10.1111/febs.13839..
Zhang X, Xu Y, He C, et al . Elevated expression of CCAT2 is associated with poor prognosis in esophageal squamous cell carcinoma[J ] . J Surg Oncol, 2015, 111(7): 834-839. DOI: 10.1002/jso.23888.
Zhang D, Cai Y, He J. Long noncoding RNA CCAT2 promotes breast tumor growth by regulating the Wnt signaling pathway[J]. Onco Targets Ther, 2015, 8: 2657-2664. DOI: 10.2147/ott.s90485.
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