• Title/Summary/Keyword: 2단계 배양방법

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Effect of Retrovirus Mediated TNF-$\alpha$ Gene Transfer to Tumor Necrosis Factor(TNF) Sensitive Tumor Cell Lines on Sensitivity to TNF (Retroviral Vector를 이용한 TNF-$\alpha$ 유전자의 이입이 암세포의 종양괴사인자(TNF) 감수성에 미치는 효과)

  • Oh, Yeon-Mok;Park, Kyeo-Yeong;Jung, Man-Pyo;Yoo, Chul-Gyu;Kim, Young-Whan;Han, Sung-Goo;Sim, Young-Soo;Han, Yong-Chol
    • Tuberculosis and Respiratory Diseases
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    • v.41 no.2
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    • pp.87-96
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    • 1994
  • Background : Since tumor necrosis factor was discovered in 1975, TNF has been well known about its cytotoxic effect on tumor cells in vivo and in vitro. According to the recent improvement of molecular biological techinques, it is possible that exogenous TNF gene is transferred to tumor cells and is expressed in theirs. By virtue of TNF gene transfer, we have expected that TNF expressed in TNF-gene-transferred tumor cells would kill tumor cells in vivo without systemic side effect. The expected mechanisms in which antitumor effects of TNF expressed in TNF-gene-transferred tumor cells are working would be as followings. In the first mechanism, TNF expressed in TNF-gene-transferred tumor cells would kill tumor cells around(like homicide). In the second mechanism, TNF expressed in TNF-gene-transferred tumor cells would kill themselves(like suicide). In the third mechanism, TNF expressed in TNF-gene-transferred tumor cells would recruit immune effector cells and kill tumor cells indirectly. In the last mechanism, TNF expressed in TNF-gene-transferred tumor cells would augment cytokine such as interferon-$\gamma$ to kill tumor cells. Among these four mechanisms of antitumor effect, only the second mechanism has not been established yet. Therefore, to elucidate the second mechanism, We performed this study. Method : We transferred TNF-$\alpha$ gene to NCI-H2058, a human mesothelioma cell line and WEHI164, a murine fibrosarcoma cell line by using retroviral vector(pLT12SNTNF). And, We determined by using MTT assay whether TNF expressed in TNF-gene-transferred tumor cell lines would kill themselves like suicide or not. Then, if TNF-gene-transferred tumor cell lines would not suicide themselves, I would know more about the TNF sensitivity of TNF-gene-transferred tumor cell lines to exogenous TNF also by MTT assay. Result : NCI-H2058 and WEHI164 which were sensitive to TNF, became far less sensitive to endogenous and exogenous TNF after being transferred TNF-$\alpha$ gene to. Conclusion : TNF-gene-transfer to NCI-H2058 and WEHI164 gave them resistance to TNF.

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Effect of Red Ginseng Total Saponin on Sciatic Nerve Regeneration (홍삼사포닌이 좌골신경 재생에 미치는 영향)

  • Han, Hye-Jeong;Lee, Hae-June;Kang, Seong-Soo;Lee, Soo-Han;Cho, Ick-Hyun;Lee, Jong-Hwan;Nah, Seung-Yeol;Park, Chang-Hyun;Uhm, Chang-Sub;Bae, Chun-Sik
    • Journal of Ginseng Research
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    • v.27 no.3
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    • pp.103-109
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    • 2003
  • We investigated the effect of ginseng total saponin (GTS) on the regeneration process of experimentally crush injured rat sciatic nerves. The bilateral sciatic nerves of fifty adult male Sprague-Dawley rats were compressed surgically with a straight hemostat for 30 seconds with 1 mm width. Twenty rats were divided into four groups to test the dose-dependent effect of GTS (0, 50, 100, or 150 mg/kg, i.p.). Saline for vehicle control group or GTS dissolved in saline was administerd for three weeks. After that period of time, the numbers of total myelinated axon and degenerated myelin in the sciatic nerves of bilateral legs were examined and analyzed using image analysis system to confirm a morphological effect of GTS. We found that the most effective concentration of GTS for the regeneration of damaged sciatic nerve was 150 mg/kg. In another set of experiment, thirty rats were divided into two groups as saline-treated vehicle group and GTS-treated group (150 mg/kg, i.p.) for three weeks. Every week we examined the numbers of total myelinated axon and degenerated myelin in the sciatic nerves of bilateral legs using image analysis system to evaluate the effect of GTS on injured nerves. We found that the regeneration of damaged sciatic nerves was facilitated in GTS-treated group compared to saline-treated group until two weeks. However, after that period of time we could not observe the significant difference between saline-treated group and GTS-treated group. These results suggest that GTS is a useful adjuvant therapy for the regeneration of the peripheral nerve injury in short period of treatment.

Experimental Studies on the Antitumor Effects of Jinryungtang Gagambang Extract (진령탕가감방의 항종양효과(抗腫瘍效果)에 관(關)한 실험적(實驗的) 연구(硏究))

  • Jeong, Jun-Tak;Moon, Goo;Moon, Suk-Jae
    • THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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    • v.4 no.1
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    • pp.37-53
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    • 1998
  • The sprig of Jinryungtang Gagambang has been used for curing as a traditional medicine without any experimental evidence to support the rational basis for their clinical use. This experiment was carried out to evaluate the possible therapeutic or antitumoral effects of Jinryungtang Gagambang extract against cancer, and to study some mechanisms responsible for its effect. The cytotoxic and antitumor effects were evaluated on human cell liens (A549, hep3B, Caki-1, Sarcoma 180) after exposure to Jinryungtang Gagambang extract using in ILS, colony forming efficency and SRB assay which were regarded as a valuable method for cytotoxic and antitumor effects of unknown compound on tumor cell lines. The results obtained in this studies were as follows. 1. As a result of exposure to Jinryungtang Gagambang extract, the proliferation of A549, hep3B, Caki-1, good correlations were shown from the results of SRB assay and those of clogenetic assay. 2. The oral administration of Jinryungtang Gagambang extract showed significant effects of increase of MST(mean survival time) and ILS(increased life span) depending on the increasing concentration. 3. Against squamous cell carcinoma induced by MCA, Jinryungtang Gagambang decreased not only the frequency of tumor production but also the number and weight of tumors per tumor bearing mice(TBM). Jinryungtang Gagambang also significantly suppressed the development of 3LL cell-implanted tumors by frequency and their size, and some developed tumors were regressed by the continuous treatment of Jinryungtang Gagambang extract into TBM. 4. Jinryungtang Gagambang extract also increased NK cell activities. According to the above results, it could be suggested that Jinryungtang Gagambang extract has prominent antiutmor effect.

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