• Asian Pacific Journal of Cancer Prevention
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• v.15 no.6
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• pp.2447-2451
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• 2014

Human papillomavirus (HPV) is the primary etiologic agent of cervical cancer. Consideration of safety and non human leukocyte antigen restriction, protein vaccine has become the most likely form of HPV therapeutic vaccine, although none have so far been reported as effective. Since tumor cells consistently express the two proteins E6 and E7, most therapeutic vaccines target one or both of them. In this study, we fabricated DC vaccines by transducing replication-defective recombinant adenoviruses expressing E6/E7 fusion gene of HPV-16, to investigate the lethal effects of specific cytotoxic T lymphocytes (CTL) against CaSki cells in vitro. Mouse immature dendritic cells (DC) were generated from bone marrow, and transfected with pAd-E6/E7 to prepare a DC vaccine and to induce specific CTL. The surface expression of CD40, CD68, MHC II and CD11c was assessed by flow cytometry (FCM), and the lethal effects of CTL against CaSki cells were determined by DAPI, FCM and CCK-8 methods. Immature mouse DC was successfully transfected by pAd-E6/E7 in vitro, and the transfecting efficiency was 40%-50%. A DC vaccine was successfully prepared and was used to induce specific CTL. Experimental results showed that the percentage of apoptosis and killing rate of CaSki cells were significantly increased by coculturing with the specific CTL (p <0.05). These results illustrated that a DC vaccine modified by HPV-16 E6/E7 gene can induce apoptosis of CaSki cells by inducing CTL, which may be used as a new strategy for biological treatment of cervical cancer.

• Radiation Oncology Journal
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• v.11 no.1
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• pp.17-27
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• 1993

To investigate the relationship between radiosensitivity and postirradiation recovery in human cancer cells, a study was performed using human cancer cell lines-A549, CaSki, SNU-C5 and PCI-13. For the study of radiosensitivity, single doses of 2, 4, 6, 8, 10, 12, and 14 Gy were given and for postirradiation recovery, two fractions of 4 Gy were separated with a time interval of 0, 0.5, 1, 1.5, 2, 2.5, 3, 4, 5, or 6 hours. Surviving fraction was estimated using colony forming ability. Surviving fractions at 2 Gy (SF2) were 0.496 (0.570-0.412) for A549, 0.496 (0.660-0.332) for CaSki, 0.386 (0.576-0.216) for SNU-C5, and 0.185 (0.247-0.123) for PCI-13. By statistical analysis the SF2 of PCI-13 was lower significantly than those of others (p<0.05). This difference was also observed at 4, 6 and 8 Gy dose levels. At 6 and 8 Gy the surviving fractions of SNU-C5 were also lower significantly than A549 and CaSki (p<0.05). By the analysis with linear quadratic model, the values of ${\alpha}$ for A549, CaSki, SNU-C5 and PCI-13 were 0.3016, 0.3212, 0.4327 and 0.8423, respectively, and those of ${\beta}$ were 0.02429, 0.02009, 0.03349 and 0.00059, respectively. So, the value of ${\alpha}$ showed increasing tendency with decreasing SF2. By the multitarget single hit model the values of Do for A549, CaSki, SUN-C5 and PCI-13 were 1.97, 1.97, 1.46 and 0.81, respectively, and those of n were 1.53, 1.50, 1.56 and 2.28, respectively. So, the value of Do decreased with decreasing SF2. Post-irradiation recovery reached plateau at around 2 hours. Recovery ratio at plateau phase ranged from 1.2 to 4.2; the value were 1.2 for PCI-13, 3.2 for CaSki, 3.3 for SNU-C5, and 4.2 for A549. Recovery rate well correlated with SF2, and increased with increasing Do and decreasing ${\alpha}$. According to above results, the intrinsic radiosensitivity was quite different among the tested cell lines; PCI-13 was the most sensitive and A549 and CaSki was similar. This difference of radiosensitivity is thought to be partly due to the difference in amount of postirradiation recovery. By linear quadratic model the difference of ${\alpha}$ values was very high, and by multitarget single hit model the difference of Do value was significantly high among four cell lines.

• Radiation Oncology Journal
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• v.17 no.3
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• pp.223-229
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• 1999

Purpose : The expression pattern of c-jun by ionizing radiation according to cell growth state (exponential growth vs. stationary phase) and its relationship with cell cycle redistribution were investigated. Materials and Methods : The exponential growth phase (day 4) and stationary phase (day 9) cells were determined from cell growth curve according to the elapse of days in CaSki. The cells were irradiated using 6 MV X-ray with a dose of 2 Gy at a fixed dose rate of 3 Gy/min. Northern blot analysis was peformed with total cellular RNA and cell cycle distribution was analyzed using flow cytometry according to time-course after irradiation. Results : The maximum expression of c-jun occurred 1 hour after irradiation in both exponential growth and stationary phase cells. After then c-jun expression was elevated upto 6 hours in exponential growth phase cells, but the level decreased in stationary phase cells. Movements of cells from G0-G1 to S, G2-M phase after irradiation were higher in exponential growth phase than stationary phase. Conclusion : c-jun may be involved in the regulation of cellular proliferation according to the growth states after irradiation.

• Korean Journal of Microbiology
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• v.28 no.2
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• pp.114-119
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• 1990

A yeast chromosomal superkiller gene (SK13) was cloned and expressed in $ski3^{-}$ Saccharomyces cerevisiae strains. The gene was fused to the structural region of E. coli lacZ gene at its C-terminus in a yeast-E. coli shuttle vector, pSR605. The fused gene complemented $ski3^{-}$ strains with SK13 activity and the quantitative level of expression was measured as determined by assaying $\beta$-galactosidase activity. The SDS-polyacrylamide gel electrophoresis and the Western blot analysis of this fused protein showed the immuno-reacted bands with a protein of the estimated molecular size (ca.250Kd).

• Proceedings of the Korean Society of Life Science Conference
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• 2008.10a
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• pp.49.2-49.2
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• 2008

• Journal of Microbiology and Biotechnology
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• v.18 no.3
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• pp.523-531
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• 2008

Radiotherapy is currently applied in the treatment of human cancers. We studied whether genistein would enhance the radiosensitivity and explored its precise molecular mechanism in cervical cancer cells. After co-treatment with genistein and irradiation, the viability, cell cycle analysis, and apoptosis signaling cascades were elucidated in CaSki cells. The viability was decreased by co-treatment with genistein and irradiation compared with irradiation treatment alone. Treatment with only ${\gamma}$-irradiation led to cell cycle arrest at the $G_1$ phase. On the other hand, co-treatment with genistein and ${\gamma}$-irradiation caused a decrease in the $G_1$ phase and a concomitant increase up to 56% in the number of $G_2$ phase. In addition, co-treatment increased the expression of p53 and p21, and Cdc2-tyr-15-p, supporting the occurrence of $G_2/M$ arrest. In general, apoptosis signaling cascades were activated by the following events: release of cytochrome c, upregulation of Bax, down regulation of Bcl-2, and activation of caspase-3 and -8 in the treatment of genistein and irradiation. Apparently, co-treatment downregulated the transcripts of E6*I, E6*II, and E7. Genistein also stimulated irradiation-induced intracellular reactive oxygene, species (ROS) production, and co-treatment-induced apoptosis was inhibited by the antioxidant N-acetylcysteine, suggesting that apoptosis has occurred through the increase in ROS by genistein and ${\gamma}$-irradiation in cervical cancer cells. Gamma-irradiation increased cyclooxygenase-1 (COX-2) expression, whereas the combination with genistein and ${\gamma}$-irradiation almost completely prevented irradiation-induced COX-2 expression and $PGE_2$ production. Co-treatment with genistein and ${\gamma}$-irradiation inhibited proliferation through $G_2/M$ arrest and induced apoptosis via ROS modulation in the CaSki cancer cells.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.1
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• pp.169-174
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• 2015

Cervical cancer (CxCa) is the most common cancer in women and a prominent cause of cancer mortality worldwide. The primary cause of CxCa is human papillomavirus (HPV). Radiation therapy and chemotherapy have been used as standard treatments, but they have undesirable side effects for patients. It was reported that gallic acid has antioxidant, antimicrobial, and anticancer activities. Gold nanoparticles are currently being used in medicine as biosensors and drug delivery agents. This study aimed to develop a drug delivery agent using gold nanoparticles conjugated with gallic acid. The study was performed in uninfected (C33A) cervical cancer cells, cervical cancer cells infected with HPV type 16 (CaSki) or 18 (HeLa), and normal Vero kidney cells. The results showed that GA inhibited the proliferation of cancer cells by inducing apoptosis. To enhance the efficacy of this anticancer activity, 15-nm spherical gold nanoparticles (GNPs) were used to deliver GA to cancer cells. The GNPs-GA complex had a reduced ability compared to unmodified GA to inhibit the growth of CxCa cells. It was interesting that high-concentration ($150{\mu}M$) GNPs-GA was not toxic to normal cells, whereas GA alone was cytotoxic. In conclusion, GNPs-GA could inhibit CxCa cell proliferation less efficiently than GA, but it was not cytotoxic to normal cells. Thus, gold nanoparticles have the potential to be used as phytochemical delivery agents for alternative cancer treatment to reduce the side effects of radiotherapy and chemotherapy.

• Journal of the Korean Society of Food Science and Nutrition
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• v.34 no.10
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• pp.1525-1530
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• 2005

The purpose of this study is to investigate anti-tumor activities, general composition, elemental composition and mineral contents of fermented liquid stem, root and fruit of Opuntia humifusa. In the general composition, the energy, crude protein, crude lipid and crude carbohydrate contents of fermented liquid stem were 86.21 Kcal, 0.92$\%$, 0.12$\%$, and 20.34$\%$, respectively. Fermented liquid fruit showed 65.32 Kcal, 1.04$\%$, 0.08$\%$, and 15.15$\%$. In mineral analysis, fermented liquid stem and fruit showed 1,800 and 388 mg of calcium per 100g. The ferrous concentrations of fermented liquid stem and fruit were 21 and 10 mg per 100 g, respectively. Methanol, ethanol and water extracts of nonfermented liquid stem and fruit did not inhibit the proliferation in human cervical cancer cells (Caski, SiHa and HaCaT), but the fermented liquid fruit showed the inhibition of Proliferation with dose-response manner in Caski and SiHa cells, but not HaCaT. Therefore, it suggests that fermented cactus may be used as one of potential adjuvant for the treatment of cervical carcinomas.

• KSBB Journal
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• v.16 no.6
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• pp.579-591
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• 2001

The cell growth inhibitor effect of cervical cancer cells was investigated by liposome mediated transfection (pRcCMVp53/lipofectin) and by transfection using adenovirus (AdCMVp57). The papilloma virus cancer cell lines we used in this study were HPV16 positive, having inhibiter gene, wild p53 gene, CaSki, SiHa, HPV18 positive HeLa, HeLaS3 and HPV negative C33A, HT3. LacZ gene of E.coli was used as the marker gene for the transfection efficiency. The effect on the inhibition of tumor cell growth was measured by cell count and cell viability though ELISA analysis and MTT assay. The inhibition of tumor cell growth was confirmed by measuring each assay for six days, comparing with the normal control cell growth. The cell growth of cervical cancer calls by transfection was significantly reduced and showed tittle differences among the cell lines. To eliminate the potential problem of Ad(adenovirus) contamination during rAAV production, rAAV can be produced by a triple transfection of vector plasmic, packaging plasmid, and adenovirus helper plasmid. To examine the helper functions of Ad plasmids on the production of rAAV vector, we carried out cotransfection of three plasmids, AAV vector, packaging construct, and Ad helper plasmids. The optimized transfection condition for calcium phosphate method is 25ug of total DNA per 10-cm-diameter plate of 293 cell. We found that rAAV yields peaked at 48hr after Ad infection. The titer of rAAV was measured by the dot blot analysis to measure the number of particles/ml based on the quantification of viral DNA. Recent1y, Kombucha(fungi) was identified as a very potent antileukefic agent. In the present study, effect of natural toxin(plankton) and Kombucha is PSP(GTXI-3, neoSTX), on various MTT assay cervical cancer cell line. Toxin(GTX 1-3, neoSTX) also inhibited the proliferation in primary cervical cancer calls in a dose-dependent toxin concentration. These results showed that toxin was very potent in inhibiting the proliferation of cervical cancer calls in vitro. Toxins and Kombuoha exhibited a dose dependent inhibition of cellular proliferation in cancer cell line.

• Biomedical Science Letters
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• v.3 no.2
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• pp.71-76
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• 1997

To develop a promising alkylating agents for anti-cervical cancer chemotherapy, five mitosene analogues were synthesized. Despite the potentiality of better cytotoxicity on solid tumor cells as opposed to that on rapidly-doubled leukemic cells, there have been no reports on the inhibition of the cervical cancer cell line by mitosene analogues. The present experiment was designed to investigate whether mitosene analogues can effectively inhibit the cellular proliferation of cervical cancer cells by using an in vitro chemosensitivty system. The mitosene analogues displayed a potent cytotoxic effect on the tested cervical cancer cell lines. Among the analogues, (22) compound gave the best inhibitory effect on SiHa tumor colonies formation. These data indicate that mitosene analogues can effectively inhibit the growth of cervical cancer cells in vitro.