LncRNA RNU12 inhibits apoptosis of hepatocellular carcinoma cells induced by recombinant human TRAIL protein through up-regulating heat shock protein 72
|更新时间:2026-01-09
|
LncRNA RNU12 inhibits apoptosis of hepatocellular carcinoma cells induced by recombinant human TRAIL protein through up-regulating heat shock protein 72
Journal of Third Military Medical UniversityVol. 39, Issue 9, Pages: 833-839(2017)
作者机构:
作者简介:
基金信息:
the General Program of National Natural Science Foundation of China(81272688)
Shihan Chen, Qingsong Deng, Jiayun Jiang, et al. LncRNA RNU12 inhibits apoptosis of hepatocellular carcinoma cells induced by recombinant human TRAIL protein through up-regulating heat shock protein 72[J]. Journal of Third Military Medical University, 2017, 39(9): 833-839.
DOI:
Shihan Chen, Qingsong Deng, Jiayun Jiang, et al. LncRNA RNU12 inhibits apoptosis of hepatocellular carcinoma cells induced by recombinant human TRAIL protein through up-regulating heat shock protein 72[J]. Journal of Third Military Medical University, 2017, 39(9): 833-839. DOI: 10.16016/j.1000-5404.201610224.
LncRNA RNU12 inhibits apoptosis of hepatocellular carcinoma cells induced by recombinant human TRAIL protein through up-regulating heat shock protein 72
摘要
Abstract
Objective
2
To investigate the molecular mechanism of inhibiting the apoptosis of hepatocellular carcinoma (HCC) cells induced by recombinant human tumor necrosis factor-related apoptosis inducing ligand (TRAIL) protein at 46~50 ℃.
Methods
2
A sub-lethal heat treatment model of HCC cells was established. The Annexin-V/PI staining was used to detect the effect of treatment of recombinant human TRAIL on the apoptotic rate of HCC cells under different temperatures (43
46 and 50 ℃
and 37 ℃ as control temperature) for 10 min. According to th
e expression profile from the Agilent human lncRNA+mRNA Array V4.0 microarray data in HCC cells in our previous study
50 ℃ specific highly expressed LncRNA RNU12 was selected as the research object. Lentiviral vector LncRNA RNU12 was constructed
and then transfected into MHCC97-H cells. The expression of heat shock protein 72 (HSP72) at mRNA and protein levels were measured in different temperature treatment groups before and after transfection by qRT-PCR and Western blotting. Meanwhile
flow cytometry was used to detect the effect of lentiviral vector LncRNA RNU12 on the apoptosis of MHCC97-H cells induced by TRAIL protein. The fluorescence
in situ
hybridization (FISH) was used to detect the sub-cellular localization of LncRNA RNU12 in HCC cells.
Results
2
Temperature at 43 ℃ promoted recombinant human TRAIL protein to induce the apoptosis in HCC cells
but it exerted opposite effect at 46~50 ℃. The mRNA level of HSP72 was significantly decreased in all 4 treatment groups after lentiviral vector infection (
P
<
0.01). And TRAIL protein levels were obviously lower at the 37
46 and 50 ℃ groups than before lentivirus-mediated interference (
P
<
0.05). After the lentiviral vector infection
the apoptosis of MHCC97-H cells induced by TRAIL protein were increased by 7.36% (
P
<
0.01) and 8.84% (
P
<
0.01)
respectively at the 46℃ and 50℃ groups. RNA-FISH showed that LncRNA RNU12 was mainly expressed in the nucleus of MHCC97-H cells at 50℃.
Conclusion
2
LncRNA RNU12 up-regulating HSP72 at transcriptional level may be one of the molecular mechanisms of inhibiting the apoptosis of HCC cells induced by recombinant human TRAIL protein at 46~50℃.
关键词
Keywords
references
Ferlay J Shin H R Bray F Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008 Int J Cancer 2010 127 12 2893 2917 10.1002/ijc.25516 Ferlay J, Shin H R, Bray F, et al. Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008[J]. Int J Cancer, 2010, 127(12): 2893-2917. DOI: 10.1002/ijc.25516.
Forner A Llovet J M Bruix J Hepatocellular carcinoma Lancet 2012 379 9822 1245 1255 10.1016/S0140-6736(11)61347-0 Forner A, Llovet J M, Bruix J. Hepatocellular carcinoma[J]. Lancet, 2012, 379(9822): 1245-1255. DOI: 10.1016/S0140-6736(11)61347-0.
Nikfarjam M Muralidharan V Christophi C Mechanisms of focal heat destruction of liver tumors J Surg Res 2005 127 2 208 223 10.1016/j.jss.2005.02.009 Nikfarjam M, Muralidharan V, Christophi C. Mechanisms of focal heat destruction of liver tumors[J]. J Surg Res, 2005, 127(2): 208-223. DOI: 10.1016/j.jss.2005.02.009.
Lim B Allen J E Prabhu V V Targeting TRAIL in the treatment of cancer: new developments Expert Opin Ther Targets 2015 19 9 1171 1185 10.1517/14728222.2015.1049838 Lim B, Allen J E, Prabhu V V, et al. Targeting TRAIL in the treatment of cancer: new developments[J]. Expert Opin Ther Targets, 2015, 19(9): 1171-1185. DOI: 10.1517/14728222.2015.1049838.
Yoo J Lee Y J Effect of hyperthermia and chemotherapeutic agents on TRAIL-induced cell death in human colon cancer cells J Cell Biochem 2008 103 1 98 109 10.1002/jcb.21389 Yoo J, Lee Y J. Effect of hyperthermia and chemotherapeutic agents on TRAIL-induced cell death in human colon cancer cells[J]. J Cell Biochem, 2008, 103(1): 98-109. DOI: 10.1002/jcb.21389.
Alcala M J Park K Yoo J Effect of hyperthermia in combination with TRAIL on the JNK-Bim signal transduction pathway and growth of xenograft tumors J Cell Biochem 2010 110 5 1073 1081 10.1002/jcb.22619 Alcala M J, Park K, Yoo J, et al. Effect of hyperthermia in combination with TRAIL on the JNK-Bim signal transduction pathway and growth of xenograft tumors[J]. J Cell Biochem, 2010, 110(5): 1073-1081. DOI: 10.1002/jcb.22619.
Song X Kim H C Kim S Y Hyperthermia-enhanced TRAIL-and mapatumumab-induced apoptotic death is mediated through mitochondria in human colon cancer cells J Cell Biochem 2012 113 5 1547 1558 10.1002/jcb.24023 Song X, Kim H C, Kim S Y, et al. Hyperthermia-enhanced TRAIL-and mapatumumab-induced apoptotic death is mediated through mitochondria in human colon cancer cells[J]. J Cell Biochem, 2012, 113(5): 1547-1558. DOI: 10.1002/jcb.24023.
Yoo J Lee Y J Effect of hyperthermia on TRAIL-induced apoptotic death in human colon cancer cells: development of a novel strategy for regional therapy J Cell Biochem 2007 101 3 619 630 10.1002/jcb.21203 Yoo J, Lee Y J. Effect of hyperthermia on TRAIL-induced apoptotic death in human colon cancer cells: development of a novel strategy for regional therapy[J]. J Cell Biochem, 2007, 101(3): 619-630. DOI: 10.1002/jcb.21203.
Wang K C Chang H Y Molecular mechanisms of long noncoding RNAs Mol Cell 2011 43 6 904 914 10.1016/j.molcel.2011.08.018 Wang K C, Chang H Y. Molecular mechanisms of long noncoding RNAs[J]. Mol Cell, 2011, 43(6): 904-914. DOI: 10.1016/j.molcel.2011.08.018.
Evans J R Feng F Y Chinnaiyan A M The bright side of dark matter: lncRNAs in cancer J Clin Invest 2016 126 8 2775 2782 10.1172/JCI84421 Evans J R, Feng F Y, Chinnaiyan A M. The bright side of dark matter: lncRNAs in cancer[J]. J Clin Invest, 2016, 126(8): 2775-2782. DOI: 10.1172/JCI84421.
Jolly C Lakhotia S C Human sat Ⅲ and Drosophila hsr omega transcripts: a common paradigm for regulation of nuclear RNA processing in stressed cells Nucleic Acids Res 2006 34 19 5508 5514 10.1093/nar/gkl711 Jolly C, Lakhotia S C. Human sat Ⅲ and Drosophila hsr omega transcripts: a common paradigm for regulation of nuclear RNA processing in stressed cells[J]. Nucleic Acids Res, 2006, 34(19): 5508-5514. DOI: 10.1093/nar/gkl711.
Lanneau D Brunet M Frisan E Heat shock proteins: essential proteins for apoptosis regulation J Cell Mol Med 2008 12 3 743 761 10.1111/j.1582-4934.2008.00273.x Lanneau D, Brunet M, Frisan E, et al. Heat shock proteins: essential proteins for apoptosis regulation[J]. J Cell Mol Med, 2008, 12(3): 743-761. DOI: 10.1111/j.1582-4934. 2008.00273.x.
Kennedy D Jager R Mosser D D Regulation of apoptosis by heat shock proteins IUBMB Life 2014 66 5 327 338 10.1002/iub.1274 Kennedy D, Jager R, Mosser D D, et al. Regulation of apoptosis by heat shock proteins[J]. IUBMB Life, 2014, 66(5): 327-338. DOI: 10.1002/iub.1274.
Taylor R P Benjamin I J Small heat shock proteins: a new classification scheme in mammals J Mol Cell Cardiol 2005 38 3 433 444 10.1016/j.yjmcc.2004.12.014 Taylor R P, Benjamin I J. Small heat shock proteins: a new classification scheme in mammals[J]. J Mol Cell Cardiol, 2005, 38(3): 433-444. DOI: 10.1016/j.yjmcc.2004.12.014.
Goloudina A R Demidov O N Garrido C Inhibition of HSP70: a challenging anti-cancer strategy Cancer Lett 2012 325 2 117 124 10.1016/j.canlet.2012.06.003 Goloudina A R, Demidov O N, Garrido C. Inhibition of HSP70: a challenging anti-cancer strategy[J]. Cancer Lett, 2012, 325(2): 117-124. DOI: 10.1016/j.canlet.2012.06.003.
Beere H M Wolf B B Cain K Heat-shock protein 70 inhibits apoptosis by preventing recruitment of procaspase-9 to the Apaf-1 apoptosome Nat Cell Biol 2000 2 8 469 475 10.1038/35019501 Beere H M, Wolf B B, Cain K, et al. Heat-shock protein 70 inhibits apoptosis by preventing recruitment of procaspase-9 to the Apaf-1 apoptosome[J]. Nat Cell Biol, 2000, 2(8): 469-475. DOI: 10.1038/35019501.
Zhang B Rong R Li H Heat shock protein 72 suppresses apoptosis by increasing the stability of X-linked inhibitor of apoptosis protein in renal ischemia/reperfusion injury Mol Med Rep 2015 11 3 1793 1799 10.3892/mmr.2014.2939 Zhang B, Rong R, Li H, et al. Heat shock protein 72 suppresses apoptosis by increasing the stability of X-linked inhibitor of apoptosis protein in renal ischemia/reperfusion injury[J]. Mol Med Rep, 2015, 11(3): 1793-1799. DOI: 10.3892/mmr.2014.2939.
Wang X Chen M Zhou J HSP27, 70 and 90, anti-apoptotic proteins, in clinical cancer therapy (Review) Int J Oncol 2014 45 1 18 30 10.3892/ijo.2014.2399 Wang X, Chen M, Zhou J, et al. HSP27, 70 and 90, anti-apoptotic proteins, in clinical cancer therapy (Review)[J]. Int J Oncol, 2014, 45(1): 18-30. DOI: 10.3892/ijo.2014.2399.