• Title/Summary/Keyword: in vitro cancer research

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Effects of $17{\beta}$-Estradiol and Estrogen Receptor Antagonists on the Proliferation of Gastric Cancer Cell Lines

  • Kim, Myung-Jin;Cho, Sung-Il;Lee, Kun-Ok;Han, Hyung-Joon;Song, Tae-Jin;Park, Seong-Heum
    • Journal of Gastric Cancer
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    • v.13 no.3
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    • pp.172-178
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    • 2013
  • Purpose: The aims of this study were as follow: 1) to de scribe the expression status of estrogen receptor-${\alpha}$ and -${\beta}$ mRNAs in five gastric carcinoma cell lines; 2) to evaluate in vitro the effects of $17{\beta}$-estradiol and estrogen receptor antagonists on the proliferation of the cell lines. Materials and Methods: Detection of estrogen receptor-${\alpha}$ and estrogen receptor-${\beta}$ mRNA in five human gastric cancer cell lines (AGS, KATO III, MKN28, MKN45 and MKN74) was made by the reverse transcription-polymerase chain reaction system. To evaluate the effect of $17{\beta}$-estradiol and estrogen receptor antagonists on the proliferation of gastric cancer cell line, the cell lines which expressed both es trogen receptors were chosen and treated with $17{\beta}$-estradiol and estrogen receptor antagonists (methyl-piperidino-pyrazole and pyrazolo [1,5-a] pyrimidine). Cell proliferation was assessed with the methylthiazol tetrazolium test. Results: Estrogen receptor-${\alpha}$ and estrogen receptor-${\beta}$ mRNAs were expressed in three (KATO III, MKN28 and MKN45) and all of the five gastric cancer cell lines, respectively. At higher concentrations, $17{\beta}$-estradiol inhibited cell growth of MKN28, MKN45 and KATO III cell lines. Neither estrogen receptor-${\alpha}$ nor estrogen receptor-${\beta}$ antagonist blocked the anti-proliferative effect of $17{\beta}$-estradiol. Conclusions: Our results indicate that estrogen receptor-${\beta}$ mRNAs are preferentially expressed in gastric cancers and also imply that hormone therapy rather than estrogen receptor blockers may be a useful strategy for the treatment of estrogen receptor-${\beta}$ positive gastric cancer. Its therapeutic significance in gastric cancer are, however, limited until more evidence of the roles of estrogen receptors in the gastric cancer are accumulated.

The Carcinogenic Liver Fluke Opisthorchis viverrini is a Reservoir for Species of Helicobacter

  • Deenonpoe, Raksawan;Chomvarin, Chariya;Pairojkul, Chawalit;Chamgramol, Yaowalux;Loukas, Alex;Brindley, Paul J;Sripa, Banchob
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.5
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    • pp.1751-1758
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    • 2015
  • There has been a strong, positive correlation between opisthorchiasis-associated cholangiocarcinoma and infection with Helicobacter. Here a rodent model of human infection with Opisthorchis viverrini was utilized to further investigate relationships of apparent co-infections with O. viverrini and H. pylori. A total of 150 hamsters were assigned to five groups: i) Control hamsters not infected with O. viverrini; ii) O. viverrini-infected hamsters; iii) non-O. viverrini infected hamsters treated with antibiotics (ABx); iv) O. viverrini-infected hamsters treated with ABx; and v) O. viverrini-infected hamsters treated both with ABx and praziquantel (PZQ). Stomach, gallbladder, liver, colonic tissue, colorectal feces and O. viverrini worms were collected and the presence of species of Helicobacter determined by PCR-based approaches. In addition, O. viverrini worms were cultured in vitro with and without ABx for four weeks, after which the presence of Helicobacter spp. was determined. In situ localization of H. pylori and Helicobacter-like species was performed using a combination of histochemistry and immunohistochemistry. The prevalence of H. pylori infection in O. viverrini-infected hamsters was significantly higher than that of O. viverrini-uninfected hamsters ($p{\leq}0.001$). Interestingly, O. viverrini-infected hamsters treated with ABx and PZQ (to remove the flukes) had a significantly lower frequency of H. pylori than either O. viverr-iniinfected hamsters treated only with ABx or O. viverrini-infected hamsters, respectively ($p{\leq}0.001$). Quantitative RT-PCR strongly confirmed the correlation between intensity H. pylori infection and the presence of liver fluke infection. In vitro, H. pylori could be detected in the O. viverrini worms cultured with ABx over four weeks. In situ localization revealed H. pylori and other Helicobacter-like bacteria in worm gut. The findings indicate that the liver fluke O. viverrini in the biliary tree of the hamsters harbors H. pylori and Helicobacter-like bacteria. Accordingly, the association between O. viverrini and H. pylori may be an obligatory mutualism.

Generation of Highly Expandable Intestinal Spheroids Composed of Stem Cells

  • Ohman Kwon;Won Dong Yu;Ye Seul Son;Kwang Bo Jung;Hana Lee;Mi-Young Son
    • International Journal of Stem Cells
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    • v.15 no.1
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    • pp.104-111
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    • 2022
  • Many of early findings regarding intestinal stem cells (ISCs) and their niche in the human intestine have relied on colorectal cancer cell lines and labor-intensive and time-consuming mouse models. However, these models cannot accurately recapitulate the physiologically relevant aspects of human ISCs. In this study, we demonstrate a reliable and robust culture method for 3D expanding intestinal spheroids (InSexp) mainly comprising ISCs and progenitors, which can be derived from 3D human intestinal organoids (HIOs). We did functional chararcterization of InSexp derived from 3D HIOs, differentiated from human pluripotent stem cells, and optimization culture methods. Our results indicate that InSexp can be rapidly expanded and easily passaged, and show enhanced growth rates via WNT pathway activation. InSexp are capable of exponential cell expansion and cryopreservation. Furthermore, in vitro-matured HIO-derived InSexp proliferate faster than immature HIO-derived InSexp with preservation of the parental HIO characteristics. These findings may facilitate the development of scalable culture systems for the long-term maintenance of human ISCs and provide an alternative platform for studying ISC biology.

Anti-Obesity Activity, Anti-Cancer Activity and Single Oral Dose Toxicity of Inonotus xeranticus Extracts (기와층버섯 추출물의 항비만활성, 항암활성 및 단회경구독성시험)

  • Kang, Eun-Hee;Lee, In-Kyoung;Hwang, Mi-Hyun;Choi, Jae-Young;Chang, Zhi-Qiang;Rhee, Man-Hee;Yun, Bong-Sik;Jiang, Cheng-Zhe;Kim, Kil-Soo;Park, Seung-Chun
    • Toxicological Research
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    • v.23 no.3
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    • pp.253-261
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    • 2007
  • In this study, we investigated the in vitro anti-obesity, anti-cancer activity and single oral dose toxicity of Inonotus xeranticus extracted by methanol (INXM) or ethyl acetate (INXE). In order to investigate anti-obestity effect of Inonotus xeranticus extracts, the 3T3-L1 cells were treated with these extracts at various concentrations(1, 10, 100 and $300{\mu}g/ml$). It was observed that 3T3-L1 cells treated with $100{\mu}g/ml$ of Inonotus obliquus ethyl acetate extract (INOE), INXM and INXE, in the absence of differentiation cocktail (0.5mM isobutylmethylxanthine (IBMX) $1{\mu}M$ dexamethasone, $1{\mu}M$ insulin), differentiated at a rate of 78.5, 80.9, and 76.4% respectively. Differentiation rates of 86.6% and 83.4% were observed in 3T3-L1 cells which were treated with differentiation cocktail at $100{\mu}g/ml$ of INXM and INXE, respectively. The anti-cancer effect of Inonotus xeranticus extracts was investigated using a method of sulforhodamine B in sarcoma 180 cell line. The cells were treated with these extracts (1, 10, 100 and $300{\mu}g/ml$) for 48 hours. The growth of cells which were treated with $300{\mu}g/ml$ of INXM was inhibited by 80.1%. The growth of sarcoma 180 cells which were treated with 100 and $300{\mu}g/ml$ of INXE was inhibited by 74.7% and 64.5%, respectively. In single oral dose toxicity study, no differences were observed between control and treated groups in clinical signs, body weight gains, and feed and water consumptions. The results indicated that Inonotus xeranticus extracts did not show any toxic effects at 2,000mg/kg in mice, and the $LD_{50}$ of these extracts was found to be higher than 2,000 mg/kg in this experiment. From the above results, Inonotus xeranticus methanol and ethyl acetate extracts might have useful clinical applications in the management of cancer and obesity and may also be useful as a medicinal food.

In Vitro and In Vivo Agreement of Microwave Radio-Thermometer and Needle Probe Thermometer During Therapeutic Ultrasound (초음파 가열 시 In Vitro 및 In Vivo에서 Microwave Radio-Thermometer와 탐침온도계의 일치도)

  • Lee, Su-Young;Cho, Sang-Hyun;Yi, Chung-Hwi;Kim, Jong-Man
    • Physical Therapy Korea
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    • v.10 no.1
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    • pp.15-27
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    • 2003
  • Therapeutic ultrasound is commonly applied for deep heating in physical therapy setting. However, it is difficult to determine the exact application dosage and to confirm the immediate heating effect. Microwave Radio-Thermometer (MRT) can measure the temperature by the electromagnetic energy in the microwave region of the object that emits above absolute zero temperature. MRT was used for early diagnosis of breast cancer since it was not harmful, non-invasive, and non-ionizing to the human body. The purposes of this study were to investigate how accurately 1.1 GHz RTM (RES Ltd. Russia) measures the change of average temperature in the tissue, and to determine the depth of temperature change measurement. Therapeutic ultrasound was applied (continuous wave for 5 minutes, 1 MHz, intensity of 1.5 $W/cm^2$ [in vitro] and 1.0 $W/cm^2$ [in vivo]) in four different conditions: (1) 30 cases of in vitro specimen of pork, (2) 30 cases of in vitro specimen of pork ankle joint, (3) 10 cases of in vivo canine thigh, and (4) 30 cases of in vivo human body. Intraclass Correlation Coeffients (ICC[3,1]) between average needle probe thermometer below surface and MRT temperature was revealed as followed: (1) Before ultrasound application ICCs ranges above .8 in specimen of pork (15 mm underneath the skin) and above .82 in specimen of pork ankle joint (10~30 mm underneath the skin). (2) After ultrasound application ICCs ranges above .7 in both specimens of pork and pork ankle joint. (3) Before ultrasound application ICCs ranges above .8 in canine thigh (20 mm underneath the skin). (4) After ultrasound application ICCs ranges above .82 in canine thigh. The temperature of the human body increased significantly with the mean of $15^{\circ}C$ in muscle tissue and with the mean of $3.5^{\circ}C$ in joint (p<.00). It was revealed that the average depth of temperature measurement of the tissue by MRT was in between 10 and 35 mm, and determined that the proper temperature measurement band was $36.5{\sim}37.0^{\circ}C$.

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Cytotoxicity of Trichothecenes to Human Solid Tumor Cells in Vitro

  • Choi, Sang-Un;Choi, Eun-Jung;Kim, Kwang-Hee;Kim, Nam-Young;Kwon, Byung-Mog;Kim, Sung-Uk;Bok, Song-Hae;Lee, So-Young;Lee, Chong-Ock
    • Archives of Pharmacal Research
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    • v.19 no.1
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    • pp.6-11
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    • 1996
  • The trichothecenes are sesquiterpenoid mycotoxins characterized by the 12,13-epoxytrichothec-9-ene ring system. We have tested cytotoxicity of several naturally-occurring or synthesized trichothecenes against human solid tumor cell lines. Among them, trichothecin(I) and $4-\beta$-Acetoxy-12,13-epoxytrichothec-9-ene (trichodermin, II) exhibited highly cytotoxic activities. 4-.betha.-Hydroxy-12,13-epoxytrichothec-9-ene (trichodermol, III) and $4-\beta$-Methoxy-12,13-epoxytrichothec-9-ene (IV) had mild cytotoxicities. But 12,13-Epoxytrichothec-9-ene-4-one (V) and $4-\beta$-Hydroxy-12,13-epoxytrichothec-9-ene(VI) had no cytotoxicities up to 10 $\mug/ml$. And in the tested cell lines, HCT15 colon cancer cell line was the most sensitive to all tested trichothecenes.

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Induction of Apoptosis and Single Strand Breaks by Extract of Pulsatilla Koreana (SB-31).

  • Kim, Sam-Yong;Kim, Hyun-Soo;Park, Sang-Jun;Kim, Jong-Suk;Park, Jee-Young;Yoon, Whan-Joong;Yoon, So-Hyun;Jo, Deog-Yeon
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1996.04a
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    • pp.174-174
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    • 1996
  • Extract of Pulsatilla Koreana (SB-31) showed promising antitumor activity in vitro (J. Kor Cancer Asso 26:959-963, 1994). We studied the mechanism of cytotoxicity of SB-31. HL-60 cells were cocultivated with various concentrations of SB-31 for 5 hours. The DNAs from HL-60 cells exposed to SB-31 showed the ladder pattern typical of apoptosis. Effect of SB-31 on topoisomerase I activity was determined by slight modification of the method by E. Aflalo(1994). The pBR322 DNA showed dose-dependent increase of R-Form DNA upon incubation with SB-31. The topoisomerase Ⅰ-like activity (Increase of R-Form DNA) was accentuated with higher dose of SB-31. It is postulated that SB-31, which is a fermentation product of Pulsatilla koreana and which loses its activity when kept in ambient temperature for more than 96 hours, may contain topoisomerase Ⅰ-like activity and the enhanced excessive single strand breaks induced by 55-31 may result in apoptosis.

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The Experimetal Effects of PalMulTang on Anti-Cancer and Immunologic Function (팔물탕(八物湯)이 항암(抗癌) 및 면역기능(免疫機能)에 미치는 실험적(實驗的) 효과(效果))

  • Park, Hae-Jun;Ko, Woo-Shin
    • The Journal of Korean Medicine
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    • v.19 no.1
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    • pp.327-338
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    • 1998
  • To investigate effect of water extract of PaIMuITang(PMT) on human cancer cell-lines and immunocytes, this research estimated proliferation of A431 cell line, KHOS-NP cell line, mouse thymocytes and mouse splenocytes, Nitric Oxide(NO) from macrophage, apoptosis and subpopulation of the mouse thymocytes. The results were obtained as follows; 1. PMT inhibited the. proliferation of A431 cell line, but it is not significant. 2. PMT inhibited the proliferation of KHOS-NP cell line, but it is not significant. 3. PMT stimulated the proliferation of mouse thymocytes, being compared Con A non-treated group. 4. PMT stimulated the proliferation of mouse splenocytes, being compared LPS treated group. 5. PMT l00g/mQ inhibited the production of NO from macrophages in vitro, being compared NPS IFN treated group. 6. PMT inhibited the production of NO from macrophages in vivo, being compared LPS|IFN treated group. 7. PMT accelerated the induction of apoptosis of the mouse thymocytes. 8. In subpopulation PMT decreased $T_H$ of the mouse thymocytes, but increased T /dT s of the mouse thymocytes.

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Synthesis and Evaluation of Antitumor Activity of 2- and 6-[(1,3- Benzothiazol-2-yl)aminomethyl]-5,8-dimethoxy-1,4-naphthoquinone Derivatives

  • Chung, Yong-Seog;Shin, Young-Kook;Zhan, Chang-Guo;Lee, Sung-Duck;Cho, Hoon
    • Archives of Pharmacal Research
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    • v.27 no.9
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    • pp.893-900
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    • 2004
  • 2- or 6-Substituted BZT-N derivatives were synthesized, and their cytotoxic activity against can-cer L1210 and SNU-1 cells was examined. The antitumor action was also assessed in mice bearing S-180 cells in peritoneal cavity. In a comparison, it was found that 6-substituted BZT-N derivatives exhibited higher potencies in both bioactivities than 2-substituted BZT-N derivatives against L1210 cells in in vitro and S-180 in vitro tests exception of compound 36. Interestingly, it was observed that 2-substituted compound 36, which has methyl group at RI position, exhib-ited a better antitumor activity than 6-substituted compounds against L1210 and SNU-1 in vitro. The EDso value of 2-substituted compound 36 against L1210 was found to be comparable to the EDso value of adriamycin and was even better against the solid cancer cell line SNU-1. It was also observed that 2-substituted compound 36 showed better antitumor activity in mice bearing S-180 cells in the peritoneal cavity. The T/C (%) value of 2-substituted compound 36 was simi-lar to that of adriamycin. Quantitative structure-activity relationship (QSAR) tests reveal that the experimental E $D_{50}$ values against SNU-1 closely correlate with both the calculated HOMO ener-gies ( $E_{HOMO}$) and the measured H-NMR chemical shift of 3-H ($\delta$$_{H}$). The results suggests that a compound having higher $E_{HOMO}$ and $\delta$$_{H}$ values usually should have a lower E $D_{50}$ (SNU-1) value.lue.lue.lue.

Radixin Knockdown by RNA Interference Suppresses Human Glioblastoma Cell Growth in Vitro and in Vivo

  • Qin, Jun-Jie;Wang, Jun-Mei;Du, Jiang;Zeng, Chun;Han, Wu;Li, Zhi-Dong;Xie, Jian;Li, Gui-Lin
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.22
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    • pp.9805-9812
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    • 2014
  • Radixin, a member of the ERM (ezrin-radixin-moesin) family, plays important roles in cell motility, invasion and tumor progression. It is expressed in a variety of normal and neoplastic cells, including many types of epithelial and lymphoid examples. However, its function in glioblastomas remains elusive. Thus, in this study, radixin gene expression was first examined in the glioblastoma cells, then suppressed with a lentivirus-mediated short-hairpin RNA (shRNA) method.We found that there were high levels of radixin expression in glioblastoma U251cells. Radixin shRNA caused down-regulation of radixin gene expression and when radixin-silenced cells were implanted into nude mice, tumor growth was significantly inhibited as compared to blank control cells or nonsense shRNA cells. In addition, microvessel density in the tumors was significantly reduced. Thrombospondin-1 (TSP-1) and E-cadherin were up-regulated in radixin- suppressed glioblastoma U251 cells. In contrast, MMP9 was down-regulated. Taken together, our findings suggest that radixin is involved in GBM cell migration and invasion, and implicate TSP-1, E-cadherin and MMP9 as metastasis-inducing factors.