• 대한임상검사과학회지
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• 제43권3호
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• pp.89-97
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• 2011

The Human Papilloma Virus (HPV) infection has been strongly associated with pathogenesis of uterine cervix carcinoma. HPV type 16, a causative agent of uterine cervix carcinoma, encodes the E6 and E7 oncogenes, expression of which is pivotal for malignant transformation and maintenance of malignant phenotypes. To develop a gene therapy for HPV-related carcinoma, We investigated the effect of E6 short-interfering RNA (E6 siRNA) on the expression of this oncogene and on the growth of HPV 16-related uterine cervix carcinoma cells. SiHa cells, a uterine cervix carcinoma cell line, which contain a single copy of HPV 16 integrated in the chromosome and express the E6 and E7 oncogenes. Before 24 hr of transfection, cells were seeded and transfected with control plasmid or E6 siRNA-expressing plasmid. The mRNA was analysed by reverse transcriptase polymerase chain reaction (RT-PCR). The cell growth rate was investigated by MTT method. The E6 mRNA level in SiHa cells was decreased in HPV 16 E6 siRNA-expression vector transfected cells and a decrease in the growth of these cells was also observed. From these results. it is evident that E6 siRNA played a role in suppression of growth of SiHa cells and has a fair chance as a candidate for gene specific therapy for HPV related uterine cervix carcinoma.

• Journal of Microbiology and Biotechnology
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• 제34권3호
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• pp.506-515
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• 2024

Primary human dermal papilla cells (HDPCs) are often preferred in studies on hair growth and regeneration. However, primary HDPCs are limited by their reduced proliferative capacity, decreased hair induction potential, and extended doubling times at higher passages. To overcome these limitations, pTARGET vectors containing human papillomavirus16 (HPV16) E6/E7 oncogenes were transfected into HDPCs and selected using G-148 to generate immortalized cells here. HPV16 E6/E7 oncogenes were efficiently transfected into primary HDPCs. Immortalized HDPC showed higher proliferative activity than primary HDPC, confirming an increased proliferation rate. Expression of p53 and pRb proteins was downregulated by E6 and E7, respectively. E6/E7 expressing HDPC cells revealed that cyclin-dependent kinase (CDK) inhibitor p21 expression was decreased, while cell cycle-related genes and proteins (CDK2 and cyclin E) and E2F family genes were upregulated. Immortalized HDPCs maintained their responsiveness to Wnt/β-catenin pathway and hair follicle formation capability, as indicated by their aggregative properties and stemness. E6/E7 immortalized HDPCs may facilitate in vitro hair growth and regeneration studies.

• E2M - 전기 전자와 첨단 소재
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• 제9권2호
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• pp.152-158
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• 1996

Optical absorption and photoconductivity spectra of undoped and Co-doped Cd$_{4}$GeSe$_{6}$ single crystals, grown by the chemical transport reaction using iodine as a transporting agent, were investigated. At 20K, the optical energy gaps of the single crystals are 1.934eV for Cd$_{4}$GeSe$_{6}$ and 1.815eV for Cd$_{4}$GeSe$_{6}$ :Co$^{2+}$. The photoconductivity spectra of these single crystals were closely investigated over the temperature range 20-290K. At 20K, the photoconductivity peaks were located at 1.797eV, 1.347eV for Cd$_{4}$GeSe$_{6}$ and 1.815eV, I,.57eV, 1.46eV and 1.38eV for Cd$_{4}$GeSe$_{6}$ :Co$^{2+}$, respectively.ely.

• Nutrition Research and Practice
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• 제3권3호
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• pp.208-211
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• 2009

This study investigated the effect of vitamin $B_6$ deficiency on antioxidant enzyme activities and lipid profile in rats with exercise-induced oxidative stress. Forty eight rats were fed either a vitamin $B_6$ deficient diet (B6-) or a control diet (control) for 4 weeks and then subdivided into 3 groups: pre-exercise (PreE); post-exercise (PostE); recess after exercise (recessE). Compared to those of control group, plasma catalase and hepatic cytosol superoxide dismutase (SOD, EC 1.15.1.1) activities of B6- group were lower regardless of exercise. The ratio of reduced glutathione/oxidized glutathione (GSH/GSSG) of B6 - group was lower in PreE and there was no difference between PostE and recessE. The level of malondialdehyde (MDA) of B6- was significantly higher in PreE and PostE. High-density lipoprotein-cholesterol (HDL-C) level of B6- group was lower regardless of exercise. Atherosclerotic index of $B_6$- group was higher in PreE and there was no difference between PostE and recessE. It is suggested that a reduction in antioxidative status caused by vitamin $B_6$ deficiency may be aggravated under exercise-induced oxidative stress.

• 대한임상검사과학회지
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• 제39권1호
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• pp.1-6
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• 2007

Cervical cancer is one of the most prevalent cancers developed in women worldwide, and human papillomavirus(HPV) type 16 is the most common agent linked to human cerivical carcinoma. Viral oncogenes E6 and E7 are selectively retained and expressed in carcinoma cells infected with human papillomavirus type 16 and cooperate with each other in the immortalization and transformation of primary keratinocytes. Because the HPV oncogenesis mechanism was not completely solved, more thorough studies are required. ln the present study, we investigated the telomere independent role of telomerase in HPV oncogenesis, we constructed the E6 mutant, E7, E6/E7 and hTERT over-expressed stable cells with a telomerase negative cell line, SW13. Expressions of inserted genes were measured by RT-PCR. E6, E7 and hTERT genes were well expressed in each cell lines when compared with the control groups. By analyzing the cell morphology under the microscope, hTERT clone size was a smaller than the mock control but oncogene expressed clones had a slightly lengthened marginal region. In addition, hTERT cells also has a tendency of brief dividing time compared to the mock control. To determine whether telomerase activity was associated with a HPV oncogenesis by oncoprotein expression, we performed the PCR based TRAP assay and a Northern blot analysis. In TRAP assay data, telomerase activities in hTERT and oncogene clones increased compared to the mock control. In addition, SW13/E6/E7 cells showed an extremely increased activity compared to the other clones. Induced hTERT mRNA by E6/E7 wasn't, however, detected in Northern blotting. In conclusion, these findings suggest that telomerase activity is closely associated with the HPV oncogenesis and E6/E7 co-expression is a most important factor of telomerase activity.

• Asian Pacific Journal of Cancer Prevention
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• 제16권9호
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• pp.3843-3847
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• 2015

Background: Cervical cancer is the second most common cause of cancer related death of women. Persistent HPV infection, especially with high-risk types such as HPV16 and HPV18, has been identified to be the primary cause of cervical cancer. E6 and E7 are the major oncoproteins of high-risk HPVs, which are expressed exclusively in HPV infected tissues, and thereby represent ideal therapeutic targets for immunotherapy of cervical cancer. Materials and Methods: In this work, we used recombinant adenovirus expressing coden-optimized HPV16 E6 and E7 fusion protein (Ad-ofE6E7) to prime dendritic cells (DC-ofE6E7), to investigate the ability of primed DC vaccine in eliciting antitumor immunity in vitro and vivo. Results: Our results indicated that DC-ofE6E7 vaccine co-culturing with splenocytes could strongly induce a tumor-specific cytotoxic T lymphocyte (CTL) response and kill the TC-1 cells effectively in vitro. Moreover, DC-ofE6E7 vaccine induced protective immunity against the challenge of TC-1 cancer cells in vivo. Conclusions: The results suggested that the HPV16 ofE6E7 primed DC vaccine has potential application for cervical cancer immunotherapy.

• Asian Pacific Journal of Cancer Prevention
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• 제15권17호
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• pp.7395-7399
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• 2014

Background: Persistent human papillomavirus (HPV) infection, especially with high-risk types such as HPV16 and HPV18, has been identified as the primary cause of cervical cancer. E6 and E7 are the major onco-proteins of high-risk HPVs, which are consistently expressed in HPV infected tissues but absent in normal tissues and represent ideal therapeutic targets for immunotherapy of cervical cancer. Materials and Methods: In this study, the optimized fusion gene HPV18 E6E7 (HPV18 ofE6E7) was constructed according to genetic codon usage for human genes. At the same time, for safety future clinical application, a mutant of HPV18 ofE6E7 fusion gene was generated by site-directed mutagenesis at L52G for the E6 protein and C98G for the E7 protein. Results: HPV18-E6E7 mutant (HPV18 ofmE6E7) constructed in this work not only lost the transformation capability for NIH 3T3 cells and tumorigenicity in BALB/c nude mice, but also maintained very good stability and antigenicity. Conclusion: These results suggest that the mutant should undergo further study for application as a safe antigenspecific therapeutic vaccine for HPV18-associated tumors.

• Tuberculosis and Respiratory Diseases
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• 제71권5호
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• pp.349-354
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• 2011

Background: Lung cancer is the leading cause of cancer deaths in the world. Human papillomavirus (HPV) infection and E6 oncoprotein expression are known risk factors for the development of non-small cell lung cancer (NSCLC). This study was performed to evaluate the prevalence of HPV 16/18 E6 oncoprotein expression in patients with NSCLC. Methods: Immunohistochemical stains of the HPV 16/18 E6 oncoprotein were performed in tumor tissues from 68 patients with NSCLC who underwent curative surgery from March 2006 to November 2008. Results: The E6 oncoprotein was expressed in 29.4% of patients with NSCLC and a statistical analysis revealed that E6 oncoprotein expression was significantly higher in females (p=0.028), never smokers (p=0.045), and patients with adenocarcinoma (p=0.022) than that in other patients. Conclusion: The E6 oncoprotein was expressed in 29.4% of patients with NSCLC. Further studies detecting HPV infection and E6 oncoprotein expression in never smoking patients with NSCLC are needed.

• Nuclear Engineering and Technology
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• 제4권3호
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• pp.214-222
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• 1972

광학급 LiF단결정의 열형광유선은 r선조사선양이 증가함에 따라서 변화한다. 즉 선량이 적을 때는 2개의 glow peak를 가지나, 선양이 정차 증가하여 10$_{5}$ rontgen 정도에 이르면 5개의 glow peak를 나타낸다. 이들 glow peak에 대응하는 energy 준위 Ei(i=1,2,3,4, 및 5)는 전도대 밑으로 부터의 깊이로 표시할 때 다음과 같은 값을 갖는다. 이들 E$_i{$의 값은 가열속도 $\theta_1$=6.6$^{\circ}C$/sec와 $\theta_2$=3.4$^{\circ}C$/sec에 대한 glow peak의 온도를 얻은 다음, Randall-Wilkins의 이론에 따라서 계산되었다. 광학급 LiF단결정에서 E$_1$과 E$_2$이외의 전자 trap은 열적으로 불안정하며 LiF(Mg) 열형광선양계에 불가결한 것으로 되어있는 sensitization의 효과가 거의 없다. LiF(Mg)는 $\theta$=6.6$^{\circ}C$/sec 일때, 17$0^{\circ}C$와 23$0^{\circ}C$에 glow peak를 나타내며 이들에 대응하는 전자 trap E$_4$와 E$_{5}$ 이외에 방사선이 조사됨에 따라서 E$_1$, E$_2$, E$_3$ 및 E$_{6}$의 전자 trap이 형성되며 이들 값은 다음과 같다. LiF(Mg)에서 방사선상해 때문에 형성된 E$_1$, E$_2$, E$_3$및 E$_{6}$는 모두 상당히 열적으로 안정하며, sensitization과정에서 형성된다. 이 안정한 6준위계에서 LiF(Mg)예 의한 방사선 선양측정이 시행되어야 한다. E$_1$,E$_2$,E$_3$ 및 E$_{6}$의 안정성은 LiF결정내의 $Mg^{$ ++/ 불순물의 영향으로 사료된다. 광학급 LiF단결정의 열형광에서 r선양의 대수표시양과 전열형광양의 대수표시 사이에 비선형성을 나타낸다. 그러나 열적으로 안정한 12$0^{\circ}C$ glow peak만을 고려하여 r선양의 대수표시양과 12$0^{\circ}C$ slow peak의 높이의 대수표시량 사이에서 비선형성이 감소되어, LiF(Mg)에 대한 곡선과 매우 유사한 곡선을 얻게 된다.

• 약학회지
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• 제48권6호
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• pp.328-334
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• 2004

Cervical cancer is one of the leading causes of female death. Viral oncoproteins E6 and E7 are selectively retained and expressed in carcinoma cells infected with HPV (Human pa pilloma virus) type 16 and cooperated in immotalization and transformation of primary keratinocyte. E6 and E7 oncoproteins interfere the functions of tumor suppressor proteins p53 and retinoblasoma protein (pRb), respectively. Among a lots of natural products, Mentha arvensis Linne var.piperascens have inhibitory effects on bindings between E6 oncoprotein and tumor suppressor p53, E3 ubiqutin- protein ligase (E6AP). HPV oncoprotein inhibitors from Mentha piperita L. were isolated by solvent partition and column chromatography (Silica gel, RP-18) and inhibitory compounds were finally purified by HPLC using an ELISA screening system based on binding between E6 and E6AP. The aim of this study is to identify the structure of inhibitory compounds and to investigate whether these compounds have inhibitory effects on functions of E6 oncoprotein. We investigated whether caffeic acid methyl ester (CAM) extracted from Mentha piperita L. could inhibit the function of E6 oncoprotein. CAM inhibited the in vitro binding of E6 and E6AP which are essential for the binding and degradation of the tumor suppressor p53 and also inhibited the proliferation of human cervical cancer cell lines (SiHa and CaSKi) in a dose response manner. These results suggest that CAM inhibited the function of E6 oncoprotein, suggesting that it can be used as a potential drug for the treatment of cervical cancers infected with HPV.