TRPV1 in Salivary Gland Epithelial Cells Is Not Involved in Salivary Secretion via Transcellular Pathway

Transient receptor potential vanilloid subtype 1 (TRPV1) was originally found in sensory neurons. Recently, it has been reported that TRPV1 is expressed in salivary gland epithelial cells (SGEC). However, the physiological role of TRPV1 in salivary secretion remains to be elucidated. We found that TRPV1 is expressed in mouse and human submandibular glands (SMG) and HSG cells, originated from human submandibular gland ducts at both mRNA and protein levels. However, capsaicin (CAP), TRPV1 agonist, had little effect on intracellular free calcium concentration ([Ca2+]i) in these cells, although carbachol consistently increased [Ca2+]i. Exposure of cells to high temperature (>43degrees C) or acidic bath solution (pH5.4) did not increase [Ca2+]i, either. We further examined the role of TRPV1 in salivary secretion using TRPV1 knock-out mice. There was no significant difference in the pilocarpine (PILO)-induced salivary flow rate between wild-type and TRPV1 knock-out mice. Saliva flow rate also showed insignificant change in the mice treated with PILO plus CAP compared with that in mice treated with PILO alone. Taken together, our results suggest that although TRPV1 is expressed in SGEC, it appears not to play any direct roles in saliva secretion via transcellular pathway.

Role of LPA and the Hippo pathway on apoptosis in salivary gland epithelial cells

Lysophosphatidic acid (LPA) is a bioactive lysophospholipid involved in numerous physiological responses. However, the expression of LPA receptors and the role of the Hippo signaling pathway in epithelial cells have remained elusive. In this experiment, we studied the functional expression of LPA receptors and the associated signaling pathway using reverse transcriptase-PCR, microspectrofluorimetry, western blotting and immunocytochemistry in salivary gland epithelial cells. We found that LPA receptors are functionally expressed and involved in activating the Hippo pathway mediated by YAP/TAZ through Lats/Mob1 and RhoA/ROCK. Upregulation of YAP/TAZ-dependent target genes, including CTGF, ANKRD1 and CYR61, has also been observed in LPA-treated cells. In addition, based on data suggesting that tumor necrosis factor (TNF)-alpha induces cell apoptosis, LPA upregulates TNF-induced caspase-3 and cleaved Poly(ADP-ribose)polymerase (PARP). However, small interfering RNA treatment to Yes-associated protein (YAP) or transcriptional co-activator with a PDZ-binding motif (TAZ) significantly decreased TNF-alpha- and LPA-induced apoptosis, suggesting that YAP and TAZ modulate the apoptotic pathway in salivary epithelial cells.

A case of ectopic pregnancy after total hysterectomy / 대한산부인과학회지

Ectopic pregnancy after total hysterectomy is a very rare event and this could be the reason for the delay in diagnosis. We report a case of a ruptured tubal pregnancy occurred after hysterectomy and diagnosed by ultrasonography, computed tomography, and laparoscopy. A 37-year-old woman visited our emergency department for evaluation of abdominal pain, 31 days after total abdominal hysterectomy. Ultrasonography and computed tomography scan showed a cystic mass on the right adnexa and a large amount of fluid collection in the pelvic cavity. The patient's urine pregnancy test was positive. Laparoscopic findings revealed a ruptured right tubal pregnancy with hemoperitoneum. Ectopic pregnancy should be included in the differential diagnosis of a patient, even after total hysterectomy has been performed.

Epithelial ovarian cancer in patients under 20 years of age / 대한산부인과학회지

OBJECTIVE: Epithelial ovarian cancer is the most common cause of death due to gynecologic malignancies in adults, but is rare in children and adolescents. This is a report of series of such patients under 20 years of age documenting their presentation, histologic type, stage of disease, treatment, and outcome. METHODS: We collected data on 21 patients with epithelial ovarian cancer under 20 years of age between January 1990 and December 2005. Patient records and pathology were reviewed. RESULTS: Epithelial ovarian cancer under 20 years of age was 2.2% in overall ovarian cancer. Epithelial ovarian cancer was 42.0% among 50 patients under 20 years of age and the most common histologic type was germ cell tumors (54%). The median age at the time of diagnosis was 17.6 years (range, 13-20 years), and the median follow-up was 87 months (range, 4-175 months). There were seventeen (81.0%) mucinous tumors, four (19.0%) serous tumors. About thirty-eight percent were low malignant potential or borderline tumors. About Eighty-five percent (18 patients) of tumors were stage I disease and about fourteen percent (3 patients) were stage III disease at the time of diagnosis. Surgical treatment included conservative surgery in 18 patients (85.7%), total abdominal hysterectomy and bilateral salpingo- oophorectomy in 3 patients (14.3%). CONCLUSION: Epithelial ovarian cancers are rare in patients in children and adolescents. The majority of ovarian cancers in this age group are mucinous tumors, stage I at diagnosis and borderline ovarian tumor. Conservative management is feasible to achieve preservation of fertility.

Value of Human Papilloma Virus load in hybrid capture II assay to predict high grade cervical intraepithelial neoplasia in menopausal women with ASCUS or LSIL / 대한산부인과학회지

OBJECTIVE: To evaluate whether determination of the Human Papilloma Virus (HPV) viral load is helpful to predict the presence of high-grade cervical intraepithelial neoplasia (CIN) that requires aggressive treatment in menopausal women who have presented with an unsatisfactory colposcopic finding and ASCUS or LSIL. METHODS: In menopausal women with ASCUS or LSIL and with who were positive for HPV, 54 women had diagnostic conization were analyzed retrospectively. We used the Hybrid Capture II method. We classified women in a low-risk group (chronic cervicitis, CIN I) and a high-risk group (> or =CIN II) according to the pathological results of conization. We also classified patients into the HPV viral load or =100 RLU/PC group. We analyzed the relationship between the pathological results of conization and viral load. RESULTS: For the conization, 30 women had chronic cervicitis, nine women had CIN I, three women had CIN II and 12 women had CIN III. In the low Hybrid Capture titer-group ( or =CIN II pathology but in the high Hybrid Capture titer-group (> or =100 RLU/PC), 11 women had high risk pathology this finding was statistically significant (P=0.013). Compared to the low Hybrid Capture titer group, the relative risk of high grade CIN (> or =CIN II) in the high Hybrid Capture titer group was 2.043 (95% CI, 1.216~3.432). CONCLUSIONS: For menopausal women with an unsatisfactory colposcopic finding and also have ASCUS or LSIL women with high HPV viral load might have tendency of > or =CIN II pathology.

The Increasing Frequency of Cervical Cancer in Korean Women under 35 / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: The goal of this study was to determine the clinical and epidemiological trends of cervical cancer in young Korean women. Social behavior including sexual habits has changed in Korean women, with sexual activity commencing at a younger age. These changes are likely to influence certain risk factors of cervical cancer, resulting in changing trends in the occurrence of the disease. MATERIALS AND METHODS: The incidence of cervical cancer in women less than 35 years-old between January 1990 and December 2006 was analyzed, and available medical records from January 1996 to December 2006 were reviewed. The clinical, pathological and epidemiologic characteristics and changing trends among these young patients were analyzed. RESULTS: Over the last two decades, the incidence of young (< 35 years) cervical cancer patients increased, more patients had an aggressive form of the disease, and there was a higher rate of women with more advanced education. Human papillomavirus (HPV) infection was detected in 94.0% of the women (63/67) tested. HPV 16 (82.5%) and HPV 18 (12.7%) were the two most common viral infections detected throughout the study period. CONCLUSIONS: The changing trends and risk factors identified suggest a need for more active education of young women about cervical cancer prevention strategies. In addition, young women are strongly recommended to undergo a regular screening test and HPV vaccination.

Adjuvant therapy in high-risk early endometrial carcinoma: a retrospective analysis of 46 cases / 부인종양

OBJECTIVE: We assessed the prognostic factors and the efficacy of adjuvant therapy and reviewed randomized studies carried out on patients receiving adjuvant therapy with early endometrial carcinoma. METHODS: One hundred and five patients that received primary surgical treatment for stage IB, IC and II endometrial cancer were enrolled in this study. The clinical outcomes were compared among the patients with variable prognostic factors and adjuvant treatments. RESULTS: One hundred and five patients fulfilled the eligibility criteria and 46 patients (43.8%) underwent adjuvant therapy. Disease recurrence occurred in nine patients within a median time of 24 months. Cervical involvement was an independent prognostic factor for the disease-free survival rates. Eight of 16 patients with FIGO stage II disease received adjuvant chemotherapy consisting of cisplatin and etoposide (or cyclophosphamide) or combined chemoradiation. The 5-year disease-free survival rate for these patients was 87.5%, a value significantly higher than for patients that received radiation therapy alone (30%). CONCLUSION: Adjuvant chemotherapy or combination chemo-radiotherapy might be superior to radiation therapy alone in high-risk early endometrial cancer patients.

Adjuvant therapy in high-risk early endometrial carcinoma: a retrospective analysis of 46 cases / 부인종양

OBJECTIVE: We assessed the prognostic factors and the efficacy of adjuvant therapy and reviewed randomized studies carried out on patients receiving adjuvant therapy with early endometrial carcinoma. METHODS: One hundred and five patients that received primary surgical treatment for stage IB, IC and II endometrial cancer were enrolled in this study. The clinical outcomes were compared among the patients with variable prognostic factors and adjuvant treatments. RESULTS: One hundred and five patients fulfilled the eligibility criteria and 46 patients (43.8%) underwent adjuvant therapy. Disease recurrence occurred in nine patients within a median time of 24 months. Cervical involvement was an independent prognostic factor for the disease-free survival rates. Eight of 16 patients with FIGO stage II disease received adjuvant chemotherapy consisting of cisplatin and etoposide (or cyclophosphamide) or combined chemoradiation. The 5-year disease-free survival rate for these patients was 87.5%, a value significantly higher than for patients that received radiation therapy alone (30%). CONCLUSION: Adjuvant chemotherapy or combination chemo-radiotherapy might be superior to radiation therapy alone in high-risk early endometrial cancer patients.

A case of mucinous borderline tumour showing persistently elevated tumor markers progressed into invasive mucinous cystadenocarcinoma / 부인종양

Mucinous tumors account for 10-15% of all epithelial of ovarian tumors, and 40% of them are borderline. Not many factors are known about progression into mucinous carcinoma of borderline ovarian tumors. The incidence of progression into invasive carcinoma is reported about 2.4% for borderline serous tumous, and 1.6% for borderline mucinous ovarian tumors. Mucinous tumors often exhibit a morphologic continuum of beningn, borderline, and invasive, so a pathologist should pay attention when examine the pathologic specimen not to miss carcinoma. This is the case of 54 female patients who developed invasive mucinous ovarian carcinoma 6 months after surgical treatment of borderline mucinous ovarian tumour.

Transcutaneous DNA immunization of HPV-16 E7 with hair-follicle specific promoter in mice / 대한산부인과학회지

OBJECTIVE: Previous reports have shown that transcutaneous immunization (TCI) with proteins or peptides in combination with adjuvants efficiently induces specific cellular and humoral immune responses. We compared the immune response after TCI with new construct which was derived from HPV-16 E7opt+K and pK6hf promoter instead of pCMV promoter and various adjuvant. METHODS: First, we made new construct ligated with HPV-16 E7 opt+K to Hair-follicle Specific pK6hf Promoter. Second, we applied pk6hf-E7 opt+K DNA with or without Lipofectamine 2000 and a combination of cholera toxin (CT) and CpG oligodeoxynucleotide (CpG) onto cold wax-depilated and hydrated bare skin of C57 BL/6 mice. To assess the ability of CTL(cytotoxic T-lymphocyte) activity, we performed intracellular cytokine staining with flow cytometric analysis to determine the number of E7-specific IFN-gamma- secreting CD8+ T cells generated in vaccinated mice with the DNA vaccine. RESULTS: Female C57BL/6 mice immunized by TCI methods with 30 microgram of pk6hf-E7 opt+K DNA with Lipofectamine2000 and CT efficiently generated E7-specific CD8(+) T cells compared with the group of pk6hf-E7 opt+K DNA only or DNA with Lipofectamine2000. CONCLUSION: Our results demonstrate that TCI of the linkaged-E7 DNA , E7 opt+K DNA to pk6hf, and Lipofectamine2000 and CT induced an antigen-specific CTL response. This result is of potential relevance for the development of therapeutic HPV-specific DNA vaccines with TCI and pK6hf promoter can be used safely.

Retraction: Comparison of As2O3 and As4O6 in the Detection of SiHa Cervical Cancer Cell Growth Inhibition Pathway / Journal of the Korean Cancer Association, 대한암학회지

No abstract available.

Prognostic outcome of patients with clinical stage I-II endometrial cancer according to bilateral salpino-oophorectomy / 대한산부인과학회지

OBJECTIVE: The aim of this study is to verify the clinical outcome of staging surgery with and (or) without bilateral salpingo-oophorectomy (BSO) in clinical stage I-II endometrial cancer patients. METHODS: We reviewed the medical records of 178 surgically treated patients in clinical stage I-II endometrial cancer between January 1994 and December 2004. Overall survival (OS) and disease free survival (DFS) were analyzed by using data gathered from the National Statistics Office. The clinical outcome was compared between patients who underwent hysterectomy with and without BSO. RESULTS: One hundred sixty patients were in clinical stage I, and 18 patients were in clinical stage II. Most of the cases showed endometrioid (93.8%) in histology and G1 (56.1%) in differentiation. BSO was performed in 142 patients. Surgico-pathological features of two group are not different but the group without BSO were younger (40.7 vs. 55.8 years old) and less myometrial invasion than the group with BSO. After mean 39.27 months follow up, we found no difference in OS and DFS between the two groups with BSO and without BSO. No factors except stage were significantly related with OS and DFS by multivariate Cox regression analysis. The rate of pelvic and paraaortic lymph node metastasis was not different between two groups. CONCLUSION: The retrospective data in the study reveals that staging surgery with and without BSO does not affect OS and DFS in clinical stage I-II endometrial cancer patients. In limited cases, such as young women, omitting BSO can be considered carefully.

The value of sentinel node HPV status for prediction of lymphnode metastasis and recurrence in cervical cancer patients / 부인종양

OBJECTIVE: The "sentinel node" is defined as the first lymph node encountered by lymphatic vessels draining a tumor. We tried to examine whether pathologic status of sentinel nodes of cervical cancer patients represent metastatic status of pelvic and paraaortic lymph nodes and to investigate the correlation between sentinel node HPV status and lymph node metastasis and recurrence of cervical cancer patients. METHODS: From August 2001 to December 2003, 57 patients affected by stage IB-IIA cervical cancer had sentinel node biopsies performed during radical hysterectomy and pelvic and paraaortic lymphadenectomy. We performed sentinel lymph node pathologic examination by frozen section and HPV typing by oligonucleotide microarray. After two years of follow up, we analyzed the prognosis of patients. RESULTS: Sentinel nodes were identified in all of our patients. A total of 79 nodes were detected as sentinel nodes. Metastasis in the sentinel nodes were found in 10 patients by frozen section and 11 patients by permanent pathologic examination. The results of sentinel lymph node frozen biopsy were statistically significant for predicting the metastatic status of the pelvic lymph nodes (p<0.05), but showed one false negative case. HPV DNA was detected in the cervices of 55 patients (96.5%), 44 (80.0%) of whom were found to have HPV DNA in the sentinel nodes. HPV DNA was detected in sentinel nodes of 10 patients among 11 patients with lymph node metastases. After mean follow up of 31.7 months (range; 1-48), disease recurred in five patients and all of these patients showed HPV in sentinel nodes. Combination of sentinel node frozen biopsy and HPV typing showed negative predictive value of 100% in predicting lymph node metastasis and recurrence. CONCLUSION: Our result suggests the possibility that sentinel node HPV typing would play a supportive role to reduce false negative rate of sentinel node biopsy. Additional study will be needed to confirm the clinical application of sentinel lymph procedure and to reveal correlation between HPV status of sentinel nodes and lymph node metastasis and recurrence of cervical cancer patients.

Clinical analysis of synchronous primary neoplasms of the female reproductive tract / 대한산부인과학회지

OBJECTIVE: A histopathologic review of synchronous primary neoplasms of the female reproductive tract was presented and the possible correlation among discrete tumor subsets, natural history, and survival was evaluated. METHODS: Between 2000 and 2005, 20 patients with synchronous primary cancers of the gynecologic malignancy were identified. Clinical and pathologic informations were obtained from medical records. Kaplan-Meier survival analysis was performed. RESULTS: The patients with Synchronous primary malignancies constituted 0.63% of all genital malignancies. The most frequently observed synchronous neoplasms were those of the ovary with the endometrium (40%). The mean age of patient with synchronous ovarian and endometrial cancer was 45.2 years. 8 patients underwent the hysterectomy with bilateral salpingo-oophorectomy and adjuvant therapy. The mean survival is 57 months (SE 10.0, 95% Confidence interval 37-77). CONCLUSION: Synchronous ovarian and endometrial cancers were detected in relatively early age and the prognosis was favorable.

Clinicopathologic characteristics and prognostic factors in adenocarcinoma of the uterine cervix / 대한산부인과학회지

OBJECTIVE: To evaluate clinicopathologic characteristics and prognostic factors in adenocarcinoma of the uterine cervix. METHODS: Medical records of patients with adenocarcinoma of the cervix who primarily underwent radical hysterectomy and pelvic lymphadenectomy (n=72) at A Hospital from August 1998 to July 2004 were retrospectively reviewed. Patients with stage IA1-IIB lesions were included in the study. Clinical and pathological data were reviewed and analyzed. RESULTS: Mean age at presentation was 46.5 years. The mean follow-up time for surviving patients was 35 months. The overall survival rate was 91.7% and disease free survival rate was 88.9%. Using univariate analysis, stage, lymph-vascular space invasion (LVSI), parametrial invasion, pelvic lymph node invasion and depth of stromal invasion were significant for survival, whereas grade, age, endometrial invasion were not significant. But using multivariate analysis, only stage and pelvic lymph node invasion remained significant independent predictors for survival. CONCLUSION: Our study demonstrates that survival rate is high for patients with early stage, after primary surgery. We found stage and pelvic lymph node invasion of significant prognostic value for survival in cervical adenocarcinoma.

Adenosine 5'-triphosphate induced NF-kappa B and intracellular calcium mobilization in cervical cancer cells / 대한산부인과학회지

OBJECTIVE: To know the effect of adenosine 5'-triphosphate (ATP) on intracellular calcium level and cell proliferation in cervical cancer cells. METHODS: Study design: Four different human cervical cancer cell lines (Caski, C33A, HeLaS3 and SiHa) were used in this study. The change of intracellular calcium level, cell proliferation and the activity of proliferation- and calcium-related transcription factors by extracellular ATP were examined in these cell lines. RESULTS: Extracellular ATP induced calcium mobilization, cell proliferation and the activation of NF-kappa B in all cell lines used. CONCLUSION: These results suggest that calcium mobilization and NF-kappa B dependent signaling pathway play an important role in the cell proliferation by ATP in cervical cancer.

The Role of Radiation Therapy for the Extramammary Paget's Disease of the Vulva ; Experience of 3 Cases / Journal of the Korean Cancer Association, 대한암학회지

We have experienced three cases of extramammary Paget's disease (EMPD) of the vulva that received radiation therapy (RT). Here, we analyze the efficacy of RT and include a literature survey. Three patients with EMPD of the vulva were treated with curative RT between 1993 and 1998. One of the patients had associated underlying adenocarcinoma of the vulva. The total doses of radiation administered were 54~78 Gy/6~8 weeks. Radiation fields encompassed 2 to 3 cm outer margins free from all visible disease including or not including the inguinal area using a 9 MeV electron or a 6 MV photon beam. Follow-up durations after radiotherapy were 0.6~11 years. Complete response was obtained in all three patients. Marginal failure occurred in one patient, and another patient with underlying adenocarcinoma treated by vulvectomy with bilateral inguinal lymph node dissection followed by external RT showed no relapse. Radiation induced side effects were transient acute confluent wet desquamation in the treated area resulting in mild late atrophic skin changes. Although surgery is currently considered the preferred primary treatment for EMPD, it has a high relapse rate due to the multifocal nature of the disease. We conclude that RT is of benefit in some selected cases of EMPD.

Proteomic analyses of cervical cancer tissues by two-dimensional gel electrophoresis and mass spectrometry / 대한산부인과학회지

OBJECTIVE: Comparison of protein expressions by two-dimensional gel electrophoresis (2-DE) in normal cervix and squamous cell carcinoma tissues in Korean women. METHODS: Normal cervix and squamous cell carcinoma tissues were solubilized with 2-DE buffer and the first dimension of PROTEAN IEF CELL, isoelectric focusing (IEF), was performed using pH3-10 linear IPG strips of 17 cm. And then running 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and sliver stain. Scanned image was analyzed using PDQuest 2-D softwareTM. Protein spot spectrum was identified by assisted laser desorption/ionization-time of fighting (MALDI-TOF) and the protein mass spectrum identifications were performed by searching protein databases of Swiss-prot/TrEMBL, Mascot and MS-FIT. RESULTS: We found 9 up-regulation proteins (Alpha enolase, Keratin 19 type I, Keratin 20 type I, Keratin 13 type I, beta-actin, Aflatoxin B1 aldehyde reductase 1, Annexin A2, Squamous cell carcinoma antigen 2, unknown), 7 down-reguation proteins (Annexin 1, Myosin regulatory light chain 2, 14-3-3 protein epsilon, Heat shock 27 kDa protein, Hypothetical protein (DKFZP434C1715), Tumor necrosis factor receptor superfamily member 13B, Smoth muscle protein 22-alpha) and 6 up and down-regulation proteins (Tropomyosin 1, Tropomyosin 2, Tropomyosin 3, Serine (or cysteine) proteinase inhibitor, Phosphatidylinositol transfer protein alpha isoform, Src homology 3 domain-containing protein HIP-55) between normal cervix and squamous cell carcinoma cell tissues. CONCLUSION: 2-DE offers total protein expressions between normal cervix and squamous cell carcinoma cell tissues, and searching of differently expressed protein for the diagnostic markers of squamous cell carcinoma tissue.

Interaction between Causative Factors of Cervical Cancer and BARD-1 (BRCA-associated ring domain-1) Polymorphism / 대한산부인과학회지

OBJECTIVE: The aim of this study was to identify interaction between causative factors of cervical cancer and BARD-1 (BRCA1 associated ring domain-1) polymorphism on cervical carcinogenesis in Korean women. METHODS: We evaluated 1,347 patients composed of women who had cervical cancer (n=337) and cervical intraepithelial neoplasia (n=308) and normal control healthy women (n=702) from October 2000 until November 2001. The single nucleotide polymorphisms (SNPs) of BARD-1 were evaluated from extracted DNA of peripheral blood with automatic DNA sequencer. The difference of each SNP between normal controls and patients was evaluated in the adjusted environmental background. RESULTS: In environmental aspect, the cervical cancer increased in the women who had the lower level of education, smoking history, the younger age of the first sexual intercourse, and the more child-bearing . The women who had BARD V507M (Arg/Arg) showed 1.6 fold increased risk in cervical cancer than the women who had BARD V507M (G/G), (G/A)(p=0.0433). And the risk increased with BARD R378S (C/C) among who have environmental risk factors compared with BARD R378S (G/G), (G/C) allele positive who have no environmental risk factors. CONCLUSION: We found that the level of education, the age at first intercourse, and the number of children were the independent risk factors in cervical carcinogenesis. The specific combination of BARD-1 and gene-environmental interactions were significantly noted in the cervical carcinogenesis in Korean women.

Transcutaneous DNA vaccination increase the CTL response: Preliminary results for the therapeutic HPV DNA vaccination / 부인종양

OBJECTIVE: Transcutaneous immunization (TCI) is a novel vaccination based on the application onto bare skin. We compared the immune response after TCI with the model DNA (OVALBUMIN) to HPV E7 and various adjuvant with intramuscular injection. We investigated the efficacy of immunization with new construct driven by K6hf promoter and compared with CMV promoter. METHODS: First, we make new construct ligated with OVA to Hair-follicle Specific pK6hf Promoter and evaluated the expression. Mouse skin was transfected with pCMV-OVA, pK6hf-OVA, pCMV-beta gal and pK6hf-beta gal and expression was determined by RT-PCR and X-Gal staining. OVA protein expression was analyzed by Western blot. Second, we immunized C57/ BL6 mice with pCMV-OVA or pK6hf-OVA DNA and cholera toxin (CT) and/or CpG. CTL was measured by ELISPOT assay of the splenocytes from the mmunized mice with the DNA vaccine. RESULTS: The beta-Galactosidase activity by X-Gal staining was detected in the epithelium of the mice skin after pK6hf-beta gal application. The mRNA and protein expression from pK6hf-OVA were evident following transcutaneous methods. Those were weaker than pCMV-OVA. TCI with pCMV-OVA and LipofectAMINE 2000 trigered an speicific CTL and Th2 response. CpG was the adjuvant for CTL after pCMV-OVA. CT and CpG did increase the CTL after pK6hf-OVA. CONCLUSION: Our data demonstrate that TCI of DNA is possible methods of CTL. CpG and CT were useful in the adjuvant for CTL. The pK6hf-OVA can induce specific CTL. This result is of potential relevance for the development of therapeutic HPV- specific DNA vaccines with TCI and pK6hf promoter can be used safely.