• Journal of Microbiology and Biotechnology
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• v.25 no.3
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• pp.413-417
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• 2015

Recently, we isolated HY253, a novel decahydrofluorene analog with a molecular structure of 7,8a-divinyl-2,4a,4b,5,6,7,8,8a,9,9a-decahydro-1H-fluorene-2,4a,4b,9a-tetraol from the roots of Aralia continentalis, which is known as Dokwhal (獨活), a traditional medicinal herb. Moreover, we previously reported its cytotoxic activity on cancer cell proliferation in human lung cancer A549 and cervical cancer HeLa cells. The current study aimed to evaluate its detailed molecular mechanisms in cell cycle arrest and apoptotic induction in human hepatocellular carcinoma HepG2 cells. Flow cytometric analysis of HepG2 cells treated with $60{\mu}M$ HY253 revealed appreciable cell cycle arrest at the G1 phase via inhibition of Rb phosphorylation and down-regulation of cyclin D1. Furthermore, using western blots, we found that up-regulation of cyclin-dependent kinase inhibitors, such as p21^CIP1 and p27^KIP1, was associated with this G₁ phase arrest. Moreover, TUNEL assay and immunoblottings revealed apoptotic induction in HepG2 cells treated with $60{\mu}M$ HY253 for 24 h, which is associated with cytochrome c release from mitochondria, via down-regulation of anti-apoptotic Bcl-2 protein, which in turn resulted in activation of caspase-9 and -3, and proteolytic cleavage of poly(ADP-ribose) polymerase (PARP). Accordingly, we suggest that HY253 may be a potent chemotherapeutic hit compound for treating human liver cancer cells via up-regulation and activation of the p53 gene.

• The Korean Journal of Cytopathology
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• v.19 no.2
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• pp.111-118
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• 2008

Background : Cervicovaginal cytology is a screening test of uterine cervical cancer. The sensitivity of cervicovaginal cytology is less than 50%, but studies of cytologic/histologic correlation are limited. We analyzed the diagnostic accuracy of cervicovaginal cytology in the detection of the squamous epithelial lesions of the uterine cervix and investigate the cause of diagnostic discordance. Materials and Methods : We collected a total of 481 sets of cervicovaginal cytology and biopsies over 5 years. The cytologic diagnoses were categorized based on The Bethesda System and the histologic diagnoses were classified as negative, flat condyloma, cervical intraepithelial neoplasia (CIN) I, CIN II, CIN III, or squamous cell carcinoma. Cytohistologic discrepancies were reviewed. Results: The concordance rate between the cytological and the histological diagnosis was 79.0%. The sensitivity and specificity of cervicovaginal cytology were 80.6% and 92.6%, respectively. Its positive predictive value and negative predictive value were 93.7% and 77.7%, respectively. The false negative rate was 19.4%. Among 54 false negative cytology cases, they were confirmed by histology as 50 flat condylomas, 2 CIN I, 1 CIN III, and 1 squamous cell carcinoma. The causes of false negative cytology were sampling errors in 75.6% and interpretation errors in 24.4%. The false positive rate was 7.4%. Among 15 false positive cytology cases, they were confirmed by histology as 12 atypical squamous cells of undetermined significance (ASCUS) and 3 low grade squamous intraepithelial lesions (LSIL). The cause of error was interpretation error in all cases. The overall diagnostic accuracy of cervicovaginal cytology was 85.7%. Conclusions : Cervicovaginal cytology shows high overall diagnostic accuracy and is a useful primary screen of uterine cervical cancer.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.7
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• pp.3367-3371
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• 2012

Objective: To investigate the effects of histone deacetylase 6 (HDAC6) siRNA on cell proliferation and cell apoptosis of the HeLa cervical carcinoma cell line and the molecular mechanisms involved. Methods: Division was into three groups: A, the untreated group; B, the control siRNA group; and C, the HDAC6 siRNA group. Lipofectamine 2000 was used for siRNA transfection, and Western blot analysis was used to determine the protein levels. Cell proliferation and apoptosis were characterized using a CCK-8 assay and flow cytometry, respectively. Results: HDAC6 protein expression in the HDAC6 siRNA-transfection group was significantly lower (P < 0.05) than in the untreated and control siRNA groups. The CCK-8 kit results demonstrated that the proliferation of HeLa cells was clearly inhibited in the HDAC6 siRNA transfection group (P < 0.05). In addition, flow cytometry revealed that the early apoptotic rate ($26.0%{\pm}0.87%$) was significantly elevated (P < 0.05) as compared with the untreated group ($10.6%{\pm}1.19%$) and control siRNA group ($8.61%{\pm}0.98%$). Furthermore, Western blot analysis indicated that bcl-2 protein expression in the HDAC6 siRNA-transfection group was down-regulated, whereas the expression of p21 and bax was up-regulated. Conclusion: HDAC6 plays an essential role in the occurrence and development of cervical carcinoma, and the down-regulation of HDAC6 expression may be useful molecular therapeutic method.

• Korean Journal of Food Science and Technology
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• v.46 no.6
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• pp.665-670
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• 2014

We isolated twelve lignans and three terpenoids were isolated from the n-hexane fraction of Schisandra chinensis extract. Using spectroscopic data and comparison with available literature, the following compounds were identified: (1) wuweizisu C, (2) gomisin N, (3) deoxyschisandrin, (4) gomisin A, (5) schisandrin, (6) chamigrenal, (7) schisanlactone D, (8) methylgomisin O, (9) angeloylgomisin O, (10) (-)-gomisin $L_2$, (11) schisandronic acid, (12) (-)-gomisin $L_1$, (13) (+)-gomisin $K_3$, (14) gomisin J, and (15) tigloylgomisin H. Notably, this was the first finding that compound (8) was isolated from this plant. Each compound was evaluated for its in vitro cytotoxic activities toward HL-60 (human leukemia), HeLa (human cervical carcinoma), and MCF-7 (breast cancer) cell lines. Compounds (7), (8), and (9) exhibited strong cytotoxic effects on HL-60 ($IC_{50}$ 7.37, 6.60, and $8.00{\mu}M$, respectively), whereas compound (6) exhibited weak cytotoxicity towards MCF-7 ($IC_{50}$ $30.50{\mu}M$). In addition, compound (8) showed the strongest activity towards HeLa cells ($IC_{50}$ $1.46{\mu}M$).

• Korean Journal of Head & Neck Oncology
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• v.18 no.1
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• pp.11-17
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• 2002

Objectives: Expression of HMGI(Y), a nucleoprotein that binds to A/T rich sequences in the minor groove of the DNA helix, is observed in neoplastically transformed cells but not in normal cells. We have analyzed HMGI(Y), p53 expression and Ki-67 labelling index in squamous cell carcinomas of the head and neck, and evaluated its clinicopathologic significance. Materials and Methods: 40 cases of squamous cell carcinoma of the head and neck were entered on the study of immunohistochemical stains for HMGI(Y), p53 and Ki-67. We analyzed the relationship between HMGI(Y), p53, Ki-67 expression and age, sex, primary tumor site, stage, survival rate, recurrence. Results: HMGI(Y) expression evidenced by immunohistochemical staining was observed in 35 of 40 (87.5%) squamous cell carcinoma of the head and neck. But no significant correlation was observed between HMGI(Y) expression and other clinical factors such as primary site, tumor stage, differenciation, cervical lymph node, metastasis, recurrence and immunohistochemical status of p53. The Ki-67 labelling index was significantly correlated with recurrence and HMGI(Y) expression (p<0.05). Conclusion: This results suggest the Ki-67 is a good prognostic factor and the HMGI(Y) expression plays some roles in carcinogenesis and cellular proliferation of squamous cell carcinoma of the head and neck. HMGI(Y) gene can be used as a cancer marker, the correlation between the gene expression and the prognosis of the cancer patient should be proved in the future studies.

• Food Science and Preservation
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• v.15 no.2
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• pp.274-279
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• 2008

This study investigated the antimutagenic and anticancer effects of soybean sauce (kanjang) supplemented with deep sea water and Sea Tangle. The Ames test indicated that kanjang had no mutagenicity but it significantly inhibited mutations induced by N-methyl-N'-nitro-N-nitrosoguanidine(MNNG) and 4-nitroquinoline-1-oxide (4NQO). Kanjang (200 ug/plate) with supplementary deep sea water and Sea Tangle had approximately 90.9% and 62.0% inhibitory effect, respectively, against mutagenesis of TA100 induced by MNNG and 4NQO. There was 61.7% inhibition of mutagenesis induced by 4NQO against the TA98 strain. Kanjang inhibited growth of cell lines of human cervical adenocarcinoma (HeLa), human hepatocellular carcinoma (Hep3B), human gastric carcinoma (AGS), human lung carcinoma (A549), and human breast adenocarcinoma (MCF-7) in a concentration-dependent manner. Treatment with kanjang supplemented with 1.0 mg/mL deep sea water had cytotoxicities of 69.4% 70.5% 55.6% 82.1 % and 73.2% against HeLa, Hep3B, AGS, A549 and MCF-7 cells respectively. In contrast kanjang supplemented with 1 mg/mL deep sea water had only $10{\sim}40%$ cytotoxicity on normal human embryonal kidney cells (293). Kanjang supplemented with deep sea water significantly inhibited tumor growth in mice injected sarcoma-180 cells. In particular, kanjang supplemented with deep sea water (25 mg/kg) inhibited tumor cell activity by 40.9%.

• Journal of the Korean Society of Food Science and Nutrition
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• v.38 no.2
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• pp.142-145
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• 2009

The cytotoxic activity against human cancer cells and anti-tumor effect in Balb/c mice of a 70% ethanol extract from masou salmon (MSE) was investigated. The cancer cell lines including human breast adenocarcinoma (MCF-7), human lung carcinoma (A549), human hepatoblastoma (HepG2), human gastric carcinoma (AGS), human cervical adenocarcinoma (HeLa) and transformed primary human embryonal kidney (293) exposed to MSE decreased cell viability as indicated by the MTT assay. The MSE shows significant cytotoxicity on MCF-7, A549, HepG2, AGS and HeLa cells, and are more active than 293 cells. The treatment with 1 mg/mL MSE resulted in 9.2%, 12.7%, 16.6%, and 16.9% cell survival against A549, MCF-7, HepG2, and AGS cells, respectively. Moreover, anticancer effect in vivo of MSE was tested in the animal system using Balb/c mice transplanted sarcoma-180 cells. MSE showed inhibition of tumor growth and the rate of inhibition was 44.7% and 55.7% at the 25 mg/kg body weight and 250 mg/kg body weight, respectively. Thus, we suggest that MSE could be a beneficial material for human cancer prevention.

• The Korean Journal of Cytopathology
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• v.11 no.1
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• pp.1-10
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• 2000

The purpose of this study is to describe the cellular characteristics of endometrial hyperplasia without/with atypia in cervical smears. These cellular features were compared with those of normal endometrium and endometrial carcinoma. We reviewed 265 cervical smears : 64 normal proliferative endometrium, 118 endometrial hyperplasia without atypia, 21 endometrial hyperplasia with atypia, and 62 endometrial adenocarcinoma. Of these smears, 72(27.2%) smears which had diagnostic endometrial epithelial cells were selected for this study. The cytologic abnormalities about cellularity, background, changes in cellular architecture, alterations in nuclear size, anisokaryosis, chromatin pattern, nucleoli, cytoplasmic vacuoles, and mitosis were observed. Nuclear enlargement(1.6 to 2 times of the nucleus in the intermediate squamous cell) and anisokaryosis(${\geq}$2 fold in size variation) were highly suggestive of endometrial hyperplasia without/with atypia. The nuclei from endometrial hyperplasia with atypia were more coarsely granular in chromatin patterns than hyperplasia without atypia(33.3% vs 3.4%). Micronucleoli were observed in all endometrial conditions, but the presence of macronucleoli were more suggestive of hyperplasia with atypia(22.2%) and adenocarcinoma(55%). The changes in cellular architecture(loss of polarity, uneven internuclear distance, overlapping and loose arrangement) were seen in hyperplasia with atypia and adenocarcinoma. Characteristically, bloody background was seen in endometrial hyperpiasia, and cellular detritus or granular proteinaceous material was only observed in endometrial adenocarcinoma. Mitoses were also observed in adenocarcinoma. In conclusion, although there is no single parameter useful for the cytologic differential diagnosis of endometrial lesions, combined cytologic evaluation can be used to diagnose hyperplasia cytologically.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.12
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• pp.5049-5053
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• 2014

Background: To investigate the expression of CD44v3 and vascular endothelial growth factor-C (VEGF-C) and their relationship with lymph node metastasis in squamous cell carcinomas (SCC) of the uterine cervix. Materials and Methods: Expression of CD44v3 and VEGF-C was analyzed in 109 cases of cervical SCC by immunohistochemistry (IHC). The relationship was analyzed between expression and the patient age, histological differentiation, formation of tumor emboli in lymphoid vessels, lymph node metastasis, FIGO staging, and TNM classification. Results: Expression rates for both CD44v3 and VEGF-C were 43.1% in cervical SCC. The cells with positive immunohistochemical staining of CD44v3 were distributed mainly around the keratin pearls in well differentiated carcinomas, but distributed diffusely in the moderately and poorly differentiated lesions. VEGF-C was found stained positively in most of the tumor cells. There were differences in expression between normal epithelium and atypical hyperplasia as well as carcinoma. Both CD44v3 and VEGF-C were found to be associated positively with lymph node metastasis and TNM classification (both p=0.000). Neither CD44v3 nor VEGF-C was found to be associated with patient age, histological differentiation, formation of tumor emboli in lymphoid vessels and FIGO staging. CD44v3 was found to be associated with VEGF-C positively (p=0.000). Conclusions: Abnormal expression of CD44v3 and VEGF-C is associated closely with the lymph node metastasis in cervical SCC, and these agents may cooperate in carcinogenesis and development of metastatic lesions.

• Biomedical Science Letters
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• v.21 no.1
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• pp.32-39
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• 2015

High-risk (HR) human papillomavirus (HPV) genotypes are strongly associated with cervical cancer, whereas other HPV genotypes are not. To identify the various HPV genotypes in clinical samples, we conducted HPV genotyping using a DNA chip test and reverse blot hybridization assay (REBA) in normal cytology samples and atypical squamous cells of undetermined significance (ASCUS) cytology samples. We also investigated the HPV infection rate and HPV genotype prevalence in women with normal cytology and ASCUS cytology. Liquid-based cytology preparations were used for the initial screening of 205 subjects with normal cytology and ASCUS cytology. The HPV infection rate was 49.8% when using the DNA chip assay and 61.0% when using the REBA test. In patients with normal cytology, the HR-HPV positive rate was 21.9% with the DNA chip assay and 43.9% with the REBA test. In contrast, 8.3% of patients with ASCUS were HR-HPV positive when using the DNA chip assay, and 13.6% were positive when tested with the REBA test. The infection rate of HR-HPV in the 40~50-year age group was significantly higher than that of the other age groups. Based on the cytological analysis of the normal and ASCUS samples, the five most prominent HPV genotypes were HPV 16, 18, 68, 33, and 58 using the DNA chip test, and they were HPV 16, 18, 53, 33, and 66 when using the REBA test. In conclusion, the findings show that the results of the REBA test are comparable to those of the DNA chip test. Most strikingly, the REBA test detected the HR-HPV genotype associated with cervical carcinoma similar to that detected with the DNA chip method. Therefore, the REBA test is a useful method to detect clinically important HR-HPV genotypes.