[The effects on expressions of p53 and PCNA by inhibition of hTERT with siRNA in transplantable model of human laryngeal squamous carcinoma in nude mice].
OBJECTIVE
To investigate the expressions of p53 and PCNA by inhibition of hTERT with siRNA in xenografted laryngeal squamous carcinoma in nude mice.


METHODS
pshRNA involved fluorescein gene was synthesized according to the sequence of hTERT cDNA. The xenografted tumor in nude mice was established by using human laryngeal squamous carcinoma cell line (Hep-2). pshRNA packaged by METAFECTENE was transfected into the transplantation tumors. The volumes of the subcutaneous tumor were determined. Fluorescence expression in the tumors was detected by confocal microscopy. The expressions of p53 and PCNA were detected using immunohistochemistry.


RESULTS
The subcutaneous tumors appeared in all the nude mice after transplantation five days later. The diameter of subcutaneous tumors become 5-7 mm after transplantation. Many green fluorescent cells were observed by confocal microscopy after transfection of pshRNA and blank plasmid. The growth of laryngeal squamous carcinoma in nude mice was significantly decreased as compared with the control group. The proliferation activity was inhibited by 76.50%. The expressions of p53 was upregulated and the expression of PCNA was downregulated.


CONCLUSIONS
The transfection of pshRNA could significantly induce inhibitory effects on growth of laryngeal squamous carcinoma in nude mice and the expression of PCNA could be effectively downregulated. The inhibition of telomerase reverse transcriptase (hTERT) mRNA could upregulate the expression of p53 and promote the apoptosis of carcinoma cells as well as inhibit the proliferation of laryngeal squamous carcinoma cells.
Zhou
Junxu
J
Department of Otolaryngology-Head and Neck Surgery, People's Hospital of Wuhan University, Wuhan, 430060, China.
Zhou
Xuhong
X
Tao
Zezhang
Z
Xiao
Bokui
B
Chen
Shiming
S
Liu
Dan
D
D000818:Animals; D002294:Carcinoma, Squamous Cell / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D045744:Cell Line, Tumor; D005260:Female; D006801:Humans; D007822:Laryngeal Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D010957:Plasmids; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism*; D034622:RNA Interference; D034741:RNA, Small Interfering; D019098:Telomerase / Q000235:genetics / Q000378:metabolism*; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism*
D018809:Proliferating Cell Nuclear Antigen; D034741:RNA, Small Interfering; C495901:TP53 protein, human; D016159:Tumor Suppressor Protein p53; D019098:Telomerase