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Cell Biology International (2010) 34, 309–315 (Printed in Great Britain)

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Overexpression of PIP5KL1 suppresses the growth of human cervical cancer cells in vitro and in vivo

Lan Shi*†‡, Kai Wang§, Mei Zhao*†‡, Xinghua Yuan*†‡ and Changzhi Huang*†‡¹

*State Key Laboratory of Molecular Oncology, Cancer Institute and Hospital, Chinese Academy of Medicine Sciences and Peking Union Medical College, Beijing 100021, Peoples Republic of China, †Department of Etiology and Carcinogenesis, Cancer Institute and Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, Peoples Republic of China, ‡Beijing Key Laboratory for Cancer Prevention, Cancer Institute and Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, Peoples Republic of China, and §Department of Obstetrics and Gynecology, Perinatal Research Laboratories, University of Wisconsin, Madison, WI 53715, USA

PIP5KL1 (phosphatidylinositol-4-phosphate 5-kinase-like 1), the fourth member of PIP5Ks (phosphatidylinositol-4-phosphate 5-kinases) type I, acts as a scaffold for localization and activation of PIP5Ks, which in turn regulate numerous cellular processes. However, the role of PIP5KL1 in the development of human cancer is poorly studied. In this study, we established a stable clone of PIP5KL1 overexpressing human cervical cancer HeLa cells. RT-PCR (reverse transcription-polymerase chain reaction) and Western immunoblot analysis were performed to testify the mRNA and protein levels of PIP5KL1 in HeLa cells. The effect of PIP5KL1 overexpression on in vitro cell growth was assessed by measuring cell proliferation and migration. The athymic nude mouse model was used to examine the effects of PIP5KL1 on tumour growth in vivo. Stable transfection of PIP5KL1 induced a significant increase in expression of both mRNA and protein levels and consequent robust inhibition of proliferation (P<0.05) and migration (P<0.05) of HeLa cells. Overexpression of PIP5KL1 significantly suppressed the growth of HeLa xenograft tumours in the flanks of nude mice. Taken together, these studies indicate a functional negative correlation between elevated levels of PIP5KL1 and the development of human cervical cancer, suggesting that PIP5KL1 overexpression may suppress cervical cancer formation.

Key words: human cervical cancer, migration, nude mice, PIP5KL1, proliferation

Abbreviations: DAPI, 4,6-diamidino-2-phenylindole, FBS, fetal bovine serum, GFP, green fluorescent protein, HPV, high-risk human papillomaviruses, MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, PIPK, phosphatidylinositol phosphate kinase, PIPKH, phosphatidylinositol phosphate kinase homolog, PIP4K, phosphatidylinositol 5-phosphate 4-kinases, PIP5K, phosphatidylinositol 4-phosphate 5-kinases, PIP5KL1, phosphatidylinositol 4-phosphate 5-kinase-like 1, PIP5Ks, phosphatidylinositol-4-phosphate 5-kinases, PI(3,4,5)P, phosphatidylinositol-3,4,5-bisphosphate, PI(4,5)P, phosphatidylinositol 4,5-bisphosphate, RPMI, Roswell Park Memorial Institute, RT-PCR, reverse transcription-polymerase chain reaction, TMA, tissue microarray

¹To whom correspondence should be addressed (email changzhihuang@yahoo.cn).

Received 14 May 2009/17 September 2009; accepted 5 November 2009

Published as Cell Biology International Immediate Publication 5 November 2009, doi:10.1042/CBI20090040

© The Author(s) Journal compilation © 2010 Portland Press Ltd