• Bulletin of the Korean Chemical Society
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• 제30권12호
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• pp.2999-3005
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• 2009

Among the more than 120 different types of human papillomavirus (HPV), types 16 and 18 have been known to be high risk agents that cause cervical cancer. We examined, in an immuno-chromatographic analysis, the potential of using the early gene product, E7 protein, as a diagnostic marker of cervical cancer caused by HPV. We developed monoclonal antibodies specific to HPV-16 and 18 E7 proteins that were produced from bacterial cells using gene recombinant technology. For each E7 protein, the optimal antibody pair was selected using the immuno-chromatographic sandwichtype binding system based on the lateral flow through membrane pores. Under these conditions, this rapid testing assay had a detection capability as low as 2 ng/mL of E7 protein. Furthermore, since viral analysis required the host cell to be lysed using chemicals such as detergents, it was possible that the E7 protein was structurally damaged during this process, which would result in a decrease in detection sensitivity. Therefore, we examined the detrimental effects caused by different detergents on the E7 protein using HeLa cells as the host. In these experiments, we found that the damage caused by the detergent, nonylphenylpolyethylene glycol (NP-40), was minimal relative to Triton X-100 commonly used for the cell lysis. Temperature also affected the stability of the E7 protein, and we found that the E7 protein was stabilized at 4$^{\circ}C$ for about 2 h, which was 4 times longer than at room temperature. Finally, a HPV-infected cervical cancer cell line, which was used as a real sample model, was treated using the optimized conditions and the presence of E7 proteins were analyzed by immuno-chromatography. The results of this experiment demonstrated that this rapid test could specifically detect HPV-infected samples.

• Asian Pacific Journal of Cancer Prevention
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• 제15권9호
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• pp.4061-4065
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• 2014

The presence of viral DNA in breast cancer cells is controversial. However, some studies have revealed a possible role for the human papillomavirus in the pathogenesis of breast cancer. The aim of the present study was to investigate the presence of HPV-DNA in breast tissue in a group of Iranian women with and without breast cancer and identification of the detected HPV types. Paraffin-embedded specimens from 65 malignant breast cancer cases and 65 cases with benign breast lesions were investigated for presence of HPV-DNA by nested polymerase chain reaction. We found HPV-DNA in 22 (33.8%) of the breast cancer specimens. All non-cancerous specimens were negative. Low and high-risk HPV types, including HPV-6 (26.2%), HPV-16 (1.5%), HPV-35 (1.5%), HPV-52 (1.5%), and HPV-11 (1.5%) were detected in our study. HPV-6 was the most prevalent type in the breast cancer specimens. Although high-risk HPV types have been shown to have a major role in cervix cancer, there have been no data that support the same relevance for other types of malignancies. Furthermore, presence of low-risk HPV types in malignancies still is a matter of debate. The data presented in this study indicates a strong need for epidemiological studies correlating different HPV types in human breast cancer.

• Asian Pacific Journal of Cancer Prevention
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• 제13권9호
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• pp.4531-4536
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• 2012

Objective: To study synergistic effects of nedaplatin and cisplatin on three human carcinoma cell lines (esophageal carcinoma cell line Eca-109, ovarian carcinoma Skov-3 and cervical carcinoma Hela). Methods: Inhibition effects were evaluated by MTT assay and cell apoptosis was detected by flow cytometry. In addition, changes of Ki-67, Bax and Bcl-2 at mRNA and protein levels were quantified by RT-PCR and Western blotting. Results: Growth inhibition in each cell lines was dose-dependent after exposure to nedaplatin or cisplatin alone. The interaction of the two drugs was synergistic at higher concentrations according to the median-effect principle. The inhibition rates with nedaplatin, cisplatin and combined treatment were $41.9{\pm}4.1%$, $47.4{\pm}2.9%$, $52.5{\pm}0.9%$(Eca-109), $39.0{\pm}1.26%$, $45.0{\pm}1.45%$, $56.2{\pm}1.44%$ (Skov-3) and $44.8{\pm}2.11%$, $46.9{\pm}0.99%$, $56.6{\pm}1.83%$ (Hela) respectively, with increase in apoptosis. Compared with the nedaplatin or cisplatin alone treatment group, the combinative treatment group's Ki-67 and bcl-2 mRNA (protein) expression was decreased while that of Bax mRNA (protein) was increased. Conclusion: Compared to the effects of nedaplatin or cisplatin alone at high concentrations, combination of nedaplatin and cisplatin at low concentrations proved to be much more effective for inhibition of proliferation and the induction of apoptosis in the Eca-109, Skov-3 and Hela cell lines.

• Asian Pacific Journal of Cancer Prevention
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• 제15권24호
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• pp.10809-10812
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• 2015

Background: Human papillomavirus (HPV) DNA testing is an effective method to screen for precancerous changes in the cervix. Samples from self-collection rather than Pap smear can potentially be used to test for HPV as they are more acceptable and preferred for use in certain settings. The objective of this study was to compare HPV DNA testing from self-collected vaginal swabs and physician-collected cervical swabs. Materials and Methods: A total of 101 self-collected vaginal and physician-collected cervical swabs of known cytology from Thai women were tested by electrochemical DNA chip assay. The specimens were divided into 4 groups: 29 with normal cytology, 14 with atypical squamous cells of undetermined significance (ASCUS), 48 with low-grade squamous intraepithelial lesion (LSIL), and 10 with high-grade squamous intraepithelial lesion (HSIL). Results: Positive detection rates of HPV from self-collected swabs were similar to those from physician-collected swabs. Among specimens with abnormal cytology, HPV was found in 50% of self-collected swabs and 47.2% of physician-collected swabs. In specimens with normal cytology, 17.2% of self-collected swabs and 24.1% of physician-collected swabs were positive for HPV. Concordance was relatively high between results from self-collected and physician-collected samples. The most common HPV genotype detected was HPV 51. Conclusions: HPV DNA testing using self-collected swabs is a feasible alternative to encourage and increase screening for cervical cancer in a population who might otherwise avoid this important preventive examination due to embarrassment, discomfort, and anxiety.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제13권4호
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• pp.391-404
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• 1991

Three anticncer agents which are different in time or dosage dependence as well as in phase specificity, namely mitomycin and adriamycin from natural products, and widely different cancer cell lines_Four epidermoid carcinomas originated from larynx, cervix, skin and gut were used toghether with one osteosarcoma as the target cell of single and combined administration of anticancer drugs. Semiautomated tetrazolium dye assay(MTT) appears to offer an attractive option for chemosensitivity of head and neck cancers since it is a simple, valid and inexpensive method of assessing chemosensitivity for large samples in a short time. The results obtained form this study were as follows. 1. Good correlations were obtained with the results of the MTT test and those of $^3H$ thymidine uptake assay. 2. $LD_{50}$ values of HIST and St.Ca. which showed relatively high doubling time on adriamycin were $30{\mu}g/ml$ and $15{\mu}g/ml$ while those of HeLa, Hep-2 and KHOS/NP were $2.1{\mu}g/ml$, $4.8{\mu}g/ml$, and $6.8{\mu}g/ml$ respectively. 3. The $LD_{50}$ value of 5-FU on five cancer cells were very high ranging from 15mg/ml to almost indefinite number, which means 5-FU is very resistant to epidermoid carcinomas or osteosarcoma examined in this study. 4. Mitomycin was relatively effective showing 80% cancer killing effect on HeLa, 70% on St. Ca. and 50% on Hep-2 at the high concentrations used. 5. Adriamycin was the most effective showing 90% cancer cell killing effect on KHOS/NP, 98% on HeLa, 80% both on Hep-2 and St. Ca. The least susceptible cancer cells toward adriamycin was HIST having only 55% cell killing effect at the high cincentration. 6. Combined therapy of adriamycin and 5-FU was more effective than single administration in all the cases examined. Most effective synergism was observed on St. Ca. at the low concentration, showing 21 times higher than each single administration.

• 대한임상검사과학회지
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• 제39권3호
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• pp.210-216
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• 2007

There is increasing evidence that stromal reaction in cancer has an important diagnostic and prognostic significance. The aim of our study is to analyze the relation between the increase in mast cell number and the expression CD34 and alpha-smooth muscle actin (${\alpha}$-SMA) in the stroma of cervical intraepithelial neoplasia (CIN) and squamous cell carcinoma (SCC). We investigated a total of 29 CIN (1,2,3) and 21 SCC (microinvasive and invasive) specimens and compared the distribution of $CD34^+$ stromal cells, ${\alpha}-SMA^+$ cells, transforming growth factor-${\beta}1$ $(TGF-{\beta}1)^+$ cells, and the density of mast cells using immunohistochemistry with antibodies against CD34, ${\alpha}$-SMA, TGF-${\beta}1$, and c-Kit (CD117) respectively. Computerized image analysis was to evaluate the positive area (%) and density of the respective immunoreactive cells. In CIN $CD34^+$ cells were abundant in the stroma but no ${\alpha}-SMA^+$ cells were identified except the wall of blood vessels. $CD34^+$ cells were progressively decreased along the continuum from CIN 2 to microinvasive SCC and not observed in the stroma of invasive SCC. Whereas ${\alpha}-SMA^+$ cells were only observed in the stroma of microinvasive and invasive SCC. We found more intense TGF-${\beta}1$ expression in the increased mast cells in the stroma of invasive SCCs than that in the stroma of CIN. These results indicate that disappearance of $CD34^+$ stromal cells and appearance of ${\alpha}-SMA^+$ cells are associated with the stromal change of CIN to SCC and the transformation of $CD34^+$ stromal cells into ${\alpha}-SMA^+$ cells is mediated by TGF-${\beta}1$ secretions in the stromal mast cell of SCC.

• 생명과학회지
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• 제32권4호
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• pp.271-278
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• 2022

신규 종양 유전자 Cancer Upregulated Gene (CUG) 2가 어떻게 유사-종양줄기세포 표현형을 유도하는지 잘 알려져 있지 않다. Cyclin E, c-Myc, Notch, 그리고 Yap1와 같은 종양단백질를 분해하여 그 발현을 조절하는 FBXW7 E3 ligase의 발현이 대장암, 자궁경부암, 그리고 위암 등 여러 암조직에서 낮아져 있음이 보고되고 있다. 그래서 우리는 이 FBXW7 단백질이 CUG2에 의한 종양형성에 관여할 수 있다는 가설을 세웠다. 이 연구에서 우리는 각 대조구 세포주보다 CUG2가 과발현된 A549 폐암 세포주와 BEAS-2B 기관지 세포주에서 FBXW7 단백질 발현이 낮게 나왔다. 여기서 MG132를 처리하게 되면 감소된 FBXW7과 FBXW7 기질로 알려진 Yap1 단백질 발현이 증가되는 결과를 관찰하였다. 종양줄기세포 현상에서 FBXW7의 역할을 규명하기 위하여, FBXW7 siRNA를 처리하였다. 대조구 세포주에서 감소된 FBXW7의 조건은 세포 이동 침습, 그리고 구형 형성이 증가되는 종양줄기세포 현상이 촉진되는 것을 관찰하였고, CUG2가 과발현된 두 세포주에서 FBXW7 발현 벡타 도입으로 FBXW7 발현 증가는 종양줄기세포 현상이 억제됨을 알 수 있었다. 또한 FBXW7의 감소는 EGFR-Akt-ERK1/2와 β-catenin-Yap1-NEK2 신호 경로를 활성화시키고, 반대로 FBXW7 발현 증가는 이 두 경로의 활성이 억제됨을 알 수 있었다. 이들 결과를 종합해 보면, CUG2 과발현은 FBXW7의 발현 감소로 이어지고, 이는 EGFR-Akt-ERK1/2와 β-catenin-Yap1-NEK2 신호경로를 활성화시켜 유사-종양줄기세포 현상을 촉진하는 것으로 생각된다.

• Asian Pacific Journal of Cancer Prevention
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• 제17권3호
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• pp.1037-1047
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• 2016

HOX transcription factors are evolutionarily conserved in many different species and are involved in important cellular processes such as morphogenesis, differentiation, and proliferation. They have also recently been implicated in carcinogenesis, but their precise role in cancer, especially in cervical cancer (CC), remains unclear. In this work, using microarray assays followed by the quantitative polymerase chain reaction (qPCR), we found that the expression of 25 HOX genes was downregulated in CC derived cell lines compared with non-tumorigenic keratinocytes. In particular, the expression of HOXA9 was observed as down-modulated in CC-derived cell lines. The expression of HOXA9 has not been previously reported in CC, or in normal keratinocytes of the cervix. We found that normal CC from women without cervical lesions express HOXA9; in contrast, CC cell lines and samples of biopsies from women with CC showed significantly diminished HOXA9 expression. Furthermore, we found that methylation at the first exon of HOXA9 could play an important role in modulating the expression of this gene. Exogenous restoration of HOXA9 expression in CC cell lines decreased cell proliferation and migration, and induced an epithelial-like phenotype. Interestingly, the silencing of human papilloma virus (HPV) E6 and E7 oncogenes induced expression of HOXA9. In conclusion, controlling HOXA9 expression appears to be a necessary step during CC development. Further studies are needed to delineate the role of HOXA9 during malignant progression and to afford more insights into the relationship between downmodulation of HOXA9 and viral HPV oncoprotein expression during cercical cancer development.

• Radiation Oncology Journal
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• 제28권2호
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• pp.91-98
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• 2010

목 적: Troglitazone (TRO)은 PPAR-$\gamma$ 작동제로서 온열감수성의 결정에 중요한 요인인 heat shock protein (HSP) 70의 합성을 저해하고 superoxide dismutase (SOD)와 카타라제를 증가시키는 것으로 알려져 있다. 이에 자궁경부암 세포를 대상으로 TRO가 온열감수성에 미치는 영향을 연구하였다. 대상 및 방법: HeLa 세포를 $5{\mu}M$ TRO로 24시간 처치한 후 $42^{\circ}C$에서 1시간 동안 온열처리를 시행하였다. 세포 생존 분획은 clonogenic assay로 측정하였다. 단백질 발현의 변화는 Western blot으로 분석하였다. SOD와 카타라제의 활성도를 측정하였으며, reactive oxygen species (ROS) 는 2',7'-dichlorofluorescin diacetate와 dihydroethidium 를 사용하여 측정하였다. 결 과: 온열처리에 의해 감소된 생존분획이 TRO 전처치에 의해 증가하였다. 온열처리에의해 HSP 70의 발현은 증가하였으나 TRO 전 처치에 의해 감소되지는 않았다. SOD와 카타라제의 활성도가 각각 1.2배, 1.3배 증가하였다. 온열처리에 의해 ROS가 증가하였으며, 증가된 ROS는 TRO 전 처치에 의해 감소하였다. 결 론: TRO는 SOD와 카타라제의 활성도를 증가시키며 이는 온열에 의한 ROS를 감소시켜 결과적으로 온열감수성을 저하시킨다.

• 대한의생명과학회지
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• 제21권1호
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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.