• The Korean Journal of Nuclear Medicine
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• v.38 no.1
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• pp.62-73
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• 2004

Purpose: The herpes simplex virus type 1 thymidine kinase gene(HSV1-tk) is an attractive candidate as a reporter gene in noninvasive reporter gene monitoring system. The HSV1-tk gene was chosen as a reporter gene, because it has been extensively studied, and there are appropriate reporter probes, substrates of HSV1-tk gene product, to apply for HSV1-tk gene imaging. We used radiolabeled 5-iodovinyl-2'-deoxyuridine (IVDU) and 5-iodovinyl-2'-fluoro-2'-deoxyuridine (IVFRU) as reporter probes for HSV1-tk gene monitoring system. Materials and Methods: We prepared HSV1-tk gene transduced Morris hepatoma cell line using retroviral vector, MOLTEN containing HSV1-tk gene. And we confirmed the HSV1-tk gene expression by Northern blotting and Western blotting. We compared in vitro uptakes of radioiodinated IVDU and IVFRU to monitor HSV1-tk gene expression in Morris hepatoma cell line (MCA) and HSV1-tk gene tranduced MCA (MCA-tk) cells until 480 minutes. We also peformed correlation analysis between percentage of HSV1-tk gene tranduced MCA cell % (MCA-tk%) and uptakes of radiolabeled IVDU or IVFRU. Results: MCA-tk cell expressed HSV1-tk mRNA and HSV1-TK protein. Two compounds showed minimal uptake in MCA, but increased uptake was observed in MCA-tk. IVDU showed 4-fold higher accumulation than IVFRU at 480 min in MCA-tk (p<0.01). Both IVDU and IVFRU uptake were linearly correlated ($R^2>0.96$) with increasing MCA-tk%. Conclusion: The radiolabeld IVDU and IVFRU showed higher specific accumulation in retrovirally HSV1-tk gene transfected Morris hepatoma cell line. Both IVDU and IVFRU could be used as good substrates for evaluation of HSV1-tk gene expression.

• Journal of the Korean Society of Food Science and Nutrition
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• v.44 no.2
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• pp.200-207
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• 2015

Various natural products or their derivatives, mostly originating from plants, fungi, and bacteria, have been exploited as therapeutic drugs to treat various human diseases. In addition to previously explored organisms, research on natural compounds has now expanded into unexamined living organisms in order to identify novel bioactive substances. Here, we determined whether or not the larval form of the mealworm beetle Tenebrio molitor, a species of darkling beetle, contains cytotoxic substances that exclusively affect cancer cell viability. Ethanol extract and its solvent partitioned fractions, hexane and ethyl acetate fractions, showed anticancer effects against various human cancer cells derived from the prostate (PC3 and 22Rv1), cervix (HeLa), liver (PLC/PRF5, HepG2, Hep3B, and SK-HEP-1), colon (HCT116), lung (NCI-H460), breast (MDA-MB231), and ovary (SKOV3). Cell death induced by the fractions was a mix of apoptosis, necrosis, and autophagy. The hexane fraction was administered intraperitoneally to nude mice bearing a hepatocellular carcinoma SK-HEP-1 and showed inhibition of tumor growth in vivo. Therefore, we concluded that worm extracts contain cytotoxic substances, which can be enriched by proper fractionation protocols, and further separation and purification could lead to the identification of novel molecules to treat human cancers.

• Nuclear Medicine and Molecular Imaging
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• v.41 no.3
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• pp.226-233
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• 2007

Purpose: Dual reporter gene imaging has several advantages for more sophisticated molecular imaging studies such as gene therapy monitoring. Herein, we have constructed hepatoma cell line expressing dual reporter genes of sodium iodide symporter (NIS) and enhanced green fluorescence protein (EGFP), and the functionalities of the genes were evaluated in vivo by nuclear and optical imaging. Materials and Methods: A pRetro-PN vector was constructed after separating NIS gene from pcDNA-NIS. RSV-EGFP-WPRE fragment separated from pLNRGW was cloned into pRetro-PN vector. The final vector expressing dual reporter genes was named pRetro-PNRGW. A human hepatoma (HepG2) cells were transfected by the retrovirus containing NIS and EGFP gene (HepG2-NE). Expression of NIS gene was confirmed by RT-PCR, radioiodine uptake and efflux studies. Expression of EGFP was confirmed by RT-PCR and fluorescence microscope. The HepG2 and HepG2-NE cells were implanted in shoulder and hindlimb of nude mice, then fluorescence image, gamma camera image and I-124 microPET image were undertaken. Results: The HepG2-NE cell was successfully constructed. RT-PCR showed NIS and EGFP mRNA expression. About 50% of cells showed fluorescence. The iodine uptake of NIS-expressed cells was about 9 times higher than control. In efflux study, $T_{1/2}$ of HepG2-NE cells was 9 min. HepG2-NE xenograft showed high signal-to-background fluorescent spots and higher iodine-uptake compared to those of HepG2 xenograft. Conclusion: A hepatoma cell line expressing NIS and EGFP dual reporter genes was successfully constructed and could be used as a potential either by therapeutic gene or imaging reporter gene.

• Journal of the Korean Society of Food Science and Nutrition
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• v.37 no.2
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• pp.148-153
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• 2008

We investigated the growth inhibitory effect of Salicornia herbacea L. (SH) on human cancer cell lines in vitro. SH was extracted with methanol (SHM), followed by further fractionation into four subfractions according to polarity: hexane (SHMH), methanol (SHMM), butanol (SHMB), and aqueous (SHMA) soluble fractions. We determined the growth inhibitory effect of these fractions against human cancer cell lines using MTT assay. Among the four subfractions of SHM, the SHMM showed the strongest cytotoxic effects on cancer cell lines. We also observed quinone reductase (QR)-inducing effect of methanol layer (SHMM) on HepG2 cells and it was determined to be 3.00 at $100\;{\mu}g/mL$ level compared to the control value of 1.0. The SHMM showed the highest induction activity of quinone reductase on HepG2 cells among the partition layers. The present work suggests that SH merits further study to confirm its chemopreventive potential.

• Applied Microscopy
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• v.39 no.4
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• pp.283-289
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• 2009

In order to examine the effect of Injinhotang extract on the liver cancer induced by N-nitrosodiethylamine (NDEA) and carbon tetrachloride ($CCl_4$) in Rats. The animals were divided into three groups. The normal (Nor) group were fed basal diet. Control (Con) group were administered with NDEA (200 mg/kgb.w., i.p.) and $CCl_4$. Injinhotang extract (IJH) group treated with Injinhotang extract (260 mg/kg/day) for 8 weeks after NDEA+$CCl_4$. Enzymic antioxidants, such as superoxide dismutase (SOD) and catalase levels were determined in all the groups of animals. The activities of SOD were significantly increased in the Con, but the activities of catalase were decreased in the Con, but the anti-oxidative enzyme activities of superoxide dismutase and catalase were increased in the IJH. In the immunohistochemistry observation, treatment of Injinhotang extract reduced the rates of p53 immunoreactivity. According to the electron microscopical observation, in the liver cancer cells were increased the smooth endoplasmic reticulum and dilated the rough endoplasmic reticulum in the Con compared with IJH. These results suggest that administration of Injinhotang extract suppress or retard NDEA and $CCl_4$-induced liver cancer.

• Radiation Oncology Journal
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• v.22 no.3
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• pp.217-224
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• 2004

Purpose: It is well known that the radiosensitivity of tumor cells can be significantly reduced under hypoxic conditions. Hypoxia-inducible factor-1 $\alpha$ (HIF-1 $\alpha$) plays a pivotal role in the essential adaptive responses to hypoxia. Therefore this study investigated the relationship between HIF-1 $\alpha$ expression and radiosensitivity. M Mouse hepatoma cell line hepafcic7 and HIF-1 $\beta$-deficient mutant cell line hepa1C4 were used to analyze the role of HIF-1 a. on radiosensitivity. These cells were exposed for 6 h to desferrioxamine (DFX) before radiation. HIF-1$\alpha$. expression was examined by Western blot. Apoptosis was assessed by DNA fragmentation, propidium iodide staining, and apoptotic cell death detection ELISA kit. Radiation sensitivity was determined using MTT assay. The radiobioiogical parameters, surviving fractions at 2 Gy and 8 Gy, and mean inactivation dose (MID) from the linear-quadratic model were used to assess radiation sensitivity in the statistical analyses. Results: The expression of HIF-1 $\alpha$. was Increased, whereas apoptosis was decreased, by radiation In the presence of DFX In hepal cl c7, but not In hepal C4. The radlosensitivity of hepal C4 cells was not significantly affected by DFX treatment. The radiosensitivlty of hepal cl c7 cells was significantly decreased in the presence of DFX Conclusion: The expression of HIF-1 w by hypoxia-mimic agent DFX reduced apoptosls and radiosensitlvity in mouse hepatoma cell line hepafclc7. These results suggested that HIF-1 u could be Induced by irradiation in hypoxic ceils of tumor masses, and that this mlght Increase radioresistance in hypoxic cells.

• Journal of Life Science
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• v.20 no.6
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• pp.877-884
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• 2010

We investigated the growth inhibitory effect of internal organs of Aplysia kurodai (AK) on proliferation in cancer cell lines in vitro. The internal organs of AK were extracted with methanol (AKM), which were then further fractionated into four subfractions by using solvent partition method, resulting in hexane (AKMH), methanol (AKMM), butanol (AKMB), and aqueous (AKMA) soluble fractions. We determined the cytotoxic effect of these four fractions in four kinds of cancer cell lines - HepG2, MCF-7, HT29 and B16-F10 - by MTT assay. Among the four subfractions of AKM, AKMM showed the strongest cytotoxic effects on all cancer cell lines which were used. Morphological changes such as membrane shrinking and blebbing of cells were also observed in AKMM treatment in HepG2 cells. In addition, we also observed quinone reductase (QR) induced effect in the methanol layer (AKMM) of HepG2 cells. AKMM showed the highest induction activity of quinone reductase on HepG2 cells among the partition layers. The QR induced effect of AKMM was determined to be 2.4 at $100\;{\mu}g/ml$ level with a control value of 1.0. Although further studies are needed, the present work suggests that internal organs of Aplysia kurodai (AK) may be a chemopreventive agent for the treatment of human cells.

• Korean Journal of Medicinal Crop Science
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• v.10 no.5
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• pp.403-408
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• 2002

We prepared extracts from bark, leaf and fruit of Sorbus commixta Hedl using by water and ethanol. To test for mutagenicity and antimutagenicity on extracts, we used Rec assay and Ames test. The result of Rec assay, the extracts of the Sorbus commixta Hedl did not show a DNA-damage activity. The results of Ames test were provided that extracts of Sorbus commixta Hedl showed $43{\sim}73\;%$ antimutagenic activities. In the inhibition ratio of the growth of several human cancer cells (A549, HepG2, MCF7), 91% of MCF7 cell growth, 94% of A549 cell growth and 91% HepG2 cell growth were inhibited by adding 1mg/ml of fruit ethanol extracts, bark ethanol extracts and fruit ethanol extracts, respectively. There was a little cytotoxicity on human normal hephatocyte (WRL68), extracts(1mg/ml) showed over the 70% survival.

• Microbiology and Biotechnology Letters
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• v.29 no.3
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• pp.181-185
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• 2001

The naturally occurring chemical indole-3-carbinol (13C), found in vegetables of the Brassica genus, is a promising anticancer agent that was shown previ- ously to induce a Gl cell cycle arrest of human breast cancer cell lines, independent of estrogen receptor signaling. The anticancer activity of 13C and the possible mechanisms of its action were explored in a human hepatocellular carcinoma cell line, HepG2. Treatment of HepG2 cells with 13C suppressed the growth of the cells. The growth sup- pression caused by 13C ($IC_{50}$/: 444$\mu$M) was found to be partially due to its ability to stop the cell cycle in HepG2 cells. Western blot analysis for the Gl phase artiest demonstrated that the expression-levels of cyclin-dependent kinase (Cdk4, Cdk6) and cyclic D were reduced strongly after treatment of Hep72 cells with 13C (4007M) for 24- 72 hrs. Furthermore, I3C selectively abolished the expression of Cdk6 in a dose- and time-dependent manner, and accordingly, inhibited the phosphorylation of retinoblastoma. Interestingly, after the HepG2 cells reached their max- imal growth arrest, the level of the p21, a well-known Cdk inhibitor, increased significantly. Therefore, it could be considered that the Gl arrest of HepG2 cells treated with 13C was due to the indirect inhibition of Cdk4/6 activities by p21 Western blot analysis for G2/M phase arrest of demonstrated the levels of Cdc2 and cyclin Bl werer reduced dramatically after the treatment of HepG2 cells with 13C ($40\mu$M) for 24-72 hrs. flow cytometry of propidium iodide-stained HepG2 cells revealed that 13C induces a Gl (53%,72hr incubation) and G2 (25%,24hr incubation) cell cycle arrest. Thus, our observations have uncovered a previously undefined antiproliferative pathway for r3C that implicates Cdk4/6 and Cdc2 as a target for cell cycle control in human HepG2 cells. However, the 13C-medi- ated cell cycle arrest and repression of Cdk4/6 production did not affect the apoptotic induction of HepG2 cell.

• Journal of the Korean Society of Food Science and Nutrition
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• v.39 no.8
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• pp.1107-1112
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• 2010

This study was conducted to investigate the effect of 70% ethanol and water extract of Korean rough rice ('Ilpum', 'Goami2', 'Keunnun', 'Sulgaeng', 'Baegjinju', and 'Heugkwang') before and after germination on proliferation of human cancer cell lines (HepG2, PC-3, and MCF-7). Antiproliferation effect was higher in ethanol extract than water extract, and was higher in after germination. 'Ilpum' ethanol extract after germination showed higher anti-proliferation effect on HepG2 and PC-3 cell lines than before germination. The cell viability on HepG2 and PC-3 cell lines of 'Ilpum' ethanol extract after germination was 27.23% and 5.05% at 1.0 mg/mL, respectively. The anti-proliferation effect on MCF-7 was the highest in 'Ilpum' and 'Heugkwang' 70% ethanol extract after germination and those cell viabilities were 7.27% and 17.00% at 0.5 mg/mL, respectively. These results suggest that germinated rough rice might have a potentially preventive effect on human cancer cells.