Impact of E6-associated protein on the proliferation and invasion of prostate cancer cells in bone metastasis.
OBJECTIVE
To understand E6 associated protein (E6-AP)'s influence on prostate cancer cell proliferation and infiltration, thus providing the theoretical basis for developing therapeutic drugs for prostate cancer metastasis to the bone.


METHODS
Electroporation was performed to introduce linear regulatory plasmid PrevTet-off-in and conjugative plasmid PrevTRE2-flag-E6AP into prostate cancer cell line to establish wild-type E6-AP over-expressing transgenic LNCaP cell line; Western blot assay was adopted to examine expression levels of E6-AP, mammalian target of rapamycin (mTOR), protein kinase B (Akt), and phosphoinositide 3-kinase (PI3K); PI3K inhibitor LY294002 was applied to all the cells and MTT assay was used to measure cell proliferation; Matrigel invasion chamber assay was adopted to detect cancer cell migration and invasion.


RESULTS
Stably transfected LNCaP cells that over expressed E6-AP had higher expression levels of PI3K, Akt, and mTOR than control LNCaP cells; MTT assay showed that E6-AP-LNCaP cells were more responsive to the inhibitory effect of LY294002; Matrigel invasion chamber assay revealed increased cell crawling and adhesiveness of E6-AP-LNCaP cells.


CONCLUSIONS
Stable over-expression of E6-AP increases the proliferation and invasion of LNCaP cells.
Department of Orthopedics, General Hospital of Chinese PLA Beijing 100853, China ; First Department of Orthopaedics, Affiliated Hospital of Beihua University Jilin 132000, Jilin Province, China.
Liyan
L
Zhang
First Department of Orthopaedics, Affiliated Hospital of Beihua University Jilin 132000, Jilin Province, China.
Xiaoguang
X
Hu
Department of Orthopedics, General Hospital of Chinese PLA Beijing 100853, China.
Jiying
J
Chen
Department of Molecular Biology of Beihua University Jilin 132013, Jilin Province, China.
Guilian
G
Fu
D001859:Bone Neoplasms / Q000201:enzymology* / Q000235:genetics / Q000556:secondary*; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D004790:Enzyme Induction; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D008297:Male; D009361:Neoplasm Invasiveness; D058539:Phosphatidylinositol 3-Kinase / Q000378:metabolism; D000081082:Phosphoinositide-3 Kinase Inhibitors; D010766:Phosphorylation; D011471:Prostatic Neoplasms / Q000201:enzymology* / Q000235:genetics / Q000473:pathology*; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D015398:Signal Transduction; D058570:TOR Serine-Threonine Kinases / Q000378:metabolism; D013997:Time Factors; D014162:Transfection; D044767:Ubiquitin-Protein Ligases / Q000096:biosynthesis* / Q000235:genetics; D015854:Up-Regulation
D000081082:Phosphoinositide-3 Kinase Inhibitors; D047428:Protein Kinase Inhibitors; C471639:UBE3A protein, human; D044767:Ubiquitin-Protein Ligases; C546842:MTOR protein, human; D058539:Phosphatidylinositol 3-Kinase; D051057:Proto-Oncogene Proteins c-akt; D058570:TOR Serine-Threonine Kinases
E6-AP; Prostate cancer; invasion; proliferation