Study of Mechanism of Esophageal Cancer Development Through MKK6 Over-expression Regulated SIRT1 Expression Level

YANG Xiao-long, HU Kai-feng, LIU Ming-jia. et al

Abstract

Tо observe the influence of silent information regulator of transcription 1 (SIRT1) level by regulation of MKK6 over-expression in esophageal cancer cell Ecal09, and then to explore the relationship between p38-MAPK-SIRTl axis and progressing of esophageal cancer. Methods MKK6 over-expression vector was successfully constructed firstly. Then the divided Ecal09 cells were treated according to four groups; empty vector group, MKK6 over-xpression group(MKK6 group), MKK6-SIRT1 ShRNA group, MKK 6-RES group. The expression of MKK6 and endogenous SIR Г1 were tested by Western blot; cell proliferation capability was detected by MTT method; cell invasion force was observed by transwell method; and cell apoptosis was detected with flow cytometry. Results pcDNA3. 1 ( + ) /myc-His A-MKK6 over-expression vector was constructed successfully and proved by sequencing. MKK6 9 s over-expressing could reduce the expression of endogenous SIRT1. The viability of Ecal09 cells was decreased. The increasing of invasion and apoptosis was observed. Conclusion There might be the p38-MAPK-SIR7T regulation axis in Ecal09 cells and affecting on a series of physiological characteristics of Ecal09 cells.

 

Keywords: MKK6 SIRTl, Ecal09, Cell activity, Cell invasion force, Cell apoptosis, Feedback regulation axis

 

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References


Haper SJ. LoGrasso P. Signaling for survival and death in neurons; the role of stress activated kinase, JNK and p381. Cell Signal, 2001; 13(5); 299-310.

Cuenda A, Rousseau S. p38 MAP-Kinases pathway regulation, function and role in human diseases. Biochim Biophys Acta,2007; 1773(8): 1358- 1375.

Zeng L, Chen R, Liang F, et al. Silent information regulator, Sirtuinl and agerelated diseases, Geriatr Gerontol lnt,2009;9 (1): 7- 15.

Kaeberlein M, McVey M, Guarente L, elal. The SIRT2/3/4 complex and SIR2 alone promote longevity in saccharomyces cerevisiae by two different mechanisms. Genes Dev, 1999; 13 (19);2570-2580.

Hong E, Lee S, Kim J. et al. Ionizing radiation induces cellular senescence of articular chondrocytes via negative regulation of SIRT1 by p38 kinase. J Biol Chem.2010;285(2): 1283-1295.

Marampon F, Gravina GL. Scarsella L. et al. Angiotensin- converting-enzyme inhibition counteracts angiotensin Ц - mediated endothelial cell dysfunction by modulating the p38/ SirTl axis. J Hypertens,2013;31( 10): 1972-1983.

Ohsugi T, Kumasaka T, Urano T, et al. Construction of a full-length human T cell leukemia virus type 1 genome form MT-2 cells containing multiple defective proviruses using overlapping polymerase chain reaction. Anal Biochem. 2004 ; 3 (92);281 -288.

Kawata Y. Yano S, Kojima H. et al. Construction of a genomic DNA library by ТА cloning. Biotechniques, 1998; 24 (4):564-565.

Morrazzani M, de Carvalho DD, Kovacic H. Monolayer versus aggregate balance in survival process for EGF-induced apoptosis in A431 carcinoma cells; implication of R()S-p38-MAPK- integrin alpha2 betal pathway. Int J Cancer.2004 ; 110(6); 788- 799.

Ting Z. Yong Z. Jia Z. et al. j3-Elemene inhibits proliferation of human glioblastoma cells and causes cell-cycle (}0,/(}l arrest via mutually compensatory activation of MKK3 and MKK6. Int J Oncol,2011;38(2);419-426.

Ren HB. Li ZS.Xu GM, et al. Dynamic changes of mitogen- activated protein kinase signal transduction in rats with severe acute pancreatitis. Chin J Dig Dis,2004 ;5(3): 123-125.

Gills JJ, Castillo SS. Zhang C, et al. Phosphatidylinositol ether lipid analogues that inhibit АКТ also independently activate the stress kinase, p38alpha through MKK3/6- independent and dependent mechanisms. J Biol Chem.2007; 282(37):27020-27029.

Tachimori Y. Nagai Y. Kanamori N, et al. Pattern of lymph node metastases of esophageal squamous cell carcinoma based on the anatomical lymphatic drainage system. Dis Esophagus, 2011;24(1);33-38.

Eads CA, Lord RV, Wickramasinghe K, et al. Epigenetic patterns in the progression of esophageal adenocarcinoma. Cancer Res,2001 ;61(8) ;3410- 3418.


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