Despite advances in surgery, radiotherapy, and chemotherapy, the survival of patients with oral squamous cell carcinoma has not significantly improved over the past several decades. Gene therapy is currently under investigation and shows us new possibility of cancer curing method. This experiment was undergone to find out the cell growth inhibition effect and evidence of apoptosis by HCCS-1(human cervical cancer suppressor-1), one of the candidates of tumor suppressor gene, transducted to human oral cancer cell line. To determine the efficiency of the adenovirus as a gene delivery vector cell line was transducted with LacZ gene and analysed with X-gal staining. Northern blot was performed to confirm the transfection with HSCC-1 gene and cell viability was assessed by cell cytotoxicity assay using cell count kit(CCK). To show the evidence of apoptosis, DNA fragmentation assay and flow cytometry(FACS) were performed. We had successfully construct the recombinant HSCC-1 adenovirus(Ad5CMV-HCCS-1), and importation efficiency was 20% at 2 MOI(multiplicity of infection), 80% at 20 MOI. Northern blot analysis showed that a single 0.6kb mRNA transcript was expressed in Ad5CMV-HCCS-1 transducted cell lines. As a result of CCK, when comparing to control subjects, transducted group showed 50% growth inhibition. In DNA fragmentation assay, according to increasing of MOI, DNA volume was diminished. In FACS analysis, DNA distribution showed fragmentation.This results imply that HCCS-1gene has growth inhibition effect in human oral cancer cell lines through apoptosis induction.
Adenoviridae* ; Apoptosis* ; Blotting, Northern ; Carcinoma, Squamous Cell ; Cell Count ; Cell Line* ; Cell Survival ; DNA ; DNA Fragmentation ; Drug Therapy ; Genes, Tumor Suppressor ; Genes, vif ; Genetic Therapy ; Humans ; Lac Operon ; Mouth Neoplasms* ; Radiotherapy ; RNA, Messenger ; Transfection ; Uterine Cervical Neoplasms

PURPOSE: The purposes of this study are to evaluate normal MR spectroscopy (MRS) data of the cerebellum and pons, and to evaluate intra-unit and inter-unit differences of the results. MATERIALS AND METHODS: Nine normal volunteers (mean age 23 years) were studied with two 1.5 Tesla MR units (A and B units). A total of four sessions of MRS study were performed with two sessions in each MR unit in each volunteer. In each session, MRS data were obtained from bilateral cerebellar hemisphere and pons. MRS was performed using spin-echo single voxel technique with repetition time of 2000 ms, echo time of 288ms and 128 averagings. Voxel size was 2x2x1.5 cm in the cerebellum and 1.5x1.5x1.5 cm in the pons. Metabolite ratios of NAA/Cho, NAA/Cr, NAA/(Cho+Cr) and Cr/Cho (both peak heights and the peak areas) were compared among the results of four sessions of MRS for evaluation of intra-unit and inter-unit differences. RESULTS: In the cerebellum, mean ISD values of peak height NAA/Cho, NAA/Cr, NAA/(Cho+Crl) and Cr/Cho ratios obtained in the first session of A unit were 1.08+/-0.16, 1.44+/-.286, 0.61+/-.09 and 0.76+/-,13, respectively. Comparing with the results of the second session, intra-unit differences were 3-7%. In B unit intra-unit differences were 1-9% , except for 22% of Cr/Cho ratio. Inter-unit differences between A and B units were 1-26%. In the pons, mean (SD values of peak height NAA/Cho, NAA/Cr, NAA/(Cho+Crl and Cr/Cho ratios obtained in the first session of A unit were 1.51+/-.35, 3.35+/-.78, 1.02+/-.16 and 0.47+/-,14, respectively. Mean NAA/Cho, NAA/Cr and NAA/(Cho+Cr) ratios of the pons were significantly higher than those of the cerebellum. Intra-unit differences were 2-21% in A unit and 4-l8% in B unit. Inter-unit differences between A and B units were 1-l8%. The mean values obtained by using peak area were generally higher and wider in range than those of peak height in both units. CONCLUSION: Mean NAA/Cho, NAA/Cr and NAA/(Cho+Cr) ratios of the cerebellum were significantly lower than those of the pons. Intra-unit differences of most metabolites ratios were generally not significant, whereas inter-unit differences were greater than intra-unit differences in more metabolite ratios. Therefore, normal MRS data of the cerebellum and the pons should be obtained in every MR unit, before MRS applications in the diseases of the cerebellum and the pons.
Cerebellum* ; Healthy Volunteers ; Magnetic Resonance Spectroscopy ; Pons* ; Spectrum Analysis ; Volunteers