Effects of RNA-binding protein quaking 5 on proliferation and cell cycle of head and neck cancer cell lines

doi:10.3877/cma.j.issn.1674-1366.2015.01.005

Abstract

Abstract:

Objective

To study the inhibition of QKI-5 on human head and neck cancer cell lines in vitro.

Methods

Western blot and immunohistochemistry were performed to detect the expression of QKI-5 in the human tongue squamous-cell carcinoma samples. Head and neck cancer cells line PCI13 and tongue squamous cell carcinoma cells lines SCC15 and SCC25 were cultured by the density of 5 × 10⁴/ml. Western blot and RT-PCR were used to detect the expression levels of protein and mRNA of QKI-5 in the three cell lines. Using viral to mediate QKI-5 overexpressing in SCC25 cells and silencing in PCI13 cells. MTT method and flow cytometry assay were conducted to detect the proliferation and apoptosis of SCC25 cells with QKI-5 overexpressing and PCI13 cells with QKI-5 silencing. Western blot was used for the detection of cyclin D1, CDK4, P27, and PARP-cleaved in SCC25 cells with QKI-5 overexpressing and PCI13 cells with QKI-5 silencing.

Results

The RNA and protein expression levels of QKI-5 were relatively low in SCC15 and SCC25 cells, while in which the PCI13 cells was higher. Protein and RNA levels of QKI-5 increased significantly after viral mediated overexpression in SCC25 cells, and which were decreased significantly after virus mediated QKI-5 silencing in PCI13 cells. After overexpression of QKI-5 in SCC25 cells, the proliferation of SCC25 decreased, and most cells stayed in the G₀/G₁ phase, while cell apoptosis increased (P < 0.05) , the expression level of cyclin D1 and CDK4 decreased (P < 0.05) and the expression level of P27 and PAPR-cleaved increased (P < 0.05) . After QKI-5 silencing in PCI13 cells, the cell proliferation of PCI13 increased (P < 0.05) , most cells went into S phase, while apoptosis decreased, and the expression level increased in cyclin D1 and CDK4 (P < 0.05) , and decreased in P27 and PARP-cleaved (P < 0.05) .

Conclusions

QKI-5 could inhibit the proliferation of head and neck cancer cells.

Key words: QKI-5, Tumor, Head and neck cancer cell lines, Proliferation, Cell cycle

Chao Ma, Wei Yan, Longkun Fan, Yanjun Guo. Effects of RNA-binding protein quaking 5 on proliferation and cell cycle of head and neck cancer cell lines[J]. Chinese Journal of Stomatological Research(Electronic Edition), 2015, 09(01): 29-36.

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Figures/Tables 6

References 23

[1]	Bello IO, Soini Y, Salo T. Prognostic evaluation of oral tongue cancer：means，markers and perspectives（Ⅰ）[J]. Oral Oncol，2010，46(9):630-635.
[2]	Bello IO, Soini Y, Salo T. Prognostic evaluation of oral tongue cancer：means，markers and perspectives（Ⅱ）[J]. Oral Oncol，2010，46(9):636-643.
[3]	Matta A, Ralhan R. Overview of current and future biologically based targeted therapies in head and neck squamous cell carcinoma[J]. Head Neck Oncol，2009(1):6.
[4]	Yang QQ, Deng YF. Long non-coding RNAs as novel biomarkers and therapeutic targets in head and neck cancers[J]. Int J Clin Exp Pathol，2014，7(4):1286-1292.
[5]	Galbiatti AL, Padovani-Junior JA, Maníglia JV，et al. Head and neck cancer：causes，prevention and treatment[J]. Braz J Otorhinolaryngol，2013，79(2):239-247.
[6]	Psyrri A, Rampias T, Vermorken JB. The current and future impact of human papillomavirus on treatment of squamous cell carcinoma of the head and neck[J]. Ann Oncol，2014，25(11):2101-2115.
[7]	Biedermann B, Hotz HR, Ciosk R. The Quaking family of RNA-binding proteins：coordinators of the cell cycle and differentiation [J]. Cell Cycle，2010，9(10):1929-1933.
[8]	Wu J, Zhou L, Tonissen K，et al. The quaking I-5 protein（QKI-5）has a novel nuclear localization signal and shuttles between the nucleus and the cytoplasm[J]. J Biol Chem，1999，274(41):29202-29210.
[9]	Kondo T, Furuta T, Mitsunaga K，et al. Genomic organization and expression analysis of the mouse qkI locus[J]. Mamm Genome，1999，10(7):662-669.
[10]	Galarneau A, Richard S. Target RNA motif and target mRNAs of the Quaking STAR protein[J]. Nat Struct Mol Biol，2005，12(8):691-698.
[11]	Dive AM, Bodhade AS, Mishra MS，et al. Histological patterns of head and neck tumors：An insight to tumor histology[J]. J Oral Maxillofac Pathol，2014，18(1):58-68.
[12]	Schmitz S, Ang KK, Vermorken J，et al. Targeted therapies for squamous cell carcinoma of the head and neck：current knowledge and future directions[J]. Cancer Treat Rev，2014，40(3):390-404.
[13]	尚伟，陆海军，李薇，等.头颈部鳞癌发病机制与治疗研究进展[J].中国口腔颌面外科杂志，2009，7(4):290-297.
[14]	Varela-Rey M, Woodhoo A, Martinez-Chantar ML，et al. Alcohol，DNA methylation，and cancer[J]. Alcohol Res，2013，35(1):25-35.
[15]	Yang G, Lu X, Wang L，et al. E2F1 and RNA binding protein QKI comprise a negative feedback in the cell cycle regulation[J]. Cell Cycle，2011，10(16):2703-2713.
[16]	Yamamoto H, Soh JW, Shirin H，et al. Comparative effects of overexpression of p27^Kip1 and p21^Cip1/Waf1 on growth and differentiation in human colon carcinoma cells[J]. Oncogene，1999，18(1):103-115.
[17]	Quaroni A, Tian JQ, Seth P，et al. p27^Kip1 is an inducer of intestinal epithelial cell differentiation[J]. Am J Physiol Cell Physiol，2000，279(4):C1045-C1057.
[18]	Deschenes C, Vézina A, Beaulieu JF，et al. Role of p27^Kip1 in human intestinal cell differentiation[J]. Gastroenterology，2001，120(2):423-438.
[19]	Yang G, Fu H, Zhang J，et al. RNA-binding protein quaking，a critical regulator of colon epithelial differentiation and a suppressor of colon cancer[J]. Gastroenterology，2010，138(1):231-240.
[20]	Mulholland PJ, Fiegler H, Mazzanti C，et al. Genomic profiling identifies discrete deletions associated with translocations in glioblastoma multiforme[J]. Cell Cycle，2006，5(7):783-791.
[21]	Ichimura K, Mungall AJ, Fiegler H，et al. Small regions of overlapping deletions on 6q26 in human astrocytic tumours identified using chromosome 6 tile path array-CGH[J]. Oncogene，2006，25(8):1261-1271.
[22]	陆劲松，沈镇宙，吴炅，等.细胞周期蛋白D1、CDK4在乳腺癌中的表达及其临床意义[J].中国癌症杂志，2001，11(2):105-108.
[23]	韩炜，钟俊，王永峰，等. PARP抑制剂用于肿瘤治疗的研究进展[J].中国新药杂志，2011，20(12):1086-1091.

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