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Retroviral Vector를 이용한 TNF-$\alpha$ 유전자의 이입이 암세포의 종양괴사인자(TNF) 감수성에 미치는 효과
오연목,박계영,정만표,유철규,김영환,한성구,심영수,한용철,Oh, Yeon-Mok,Park, Kyeo-Yeong,Jung, Man-Pyo,Yoo, Chul-Gyu,Kim, Young-Whan,Han, Sung-Goo,Sim, Young-Soo,Han, Yong-Chol 대한결핵및호흡기학회 1994 Tuberculosis and Respiratory Diseases Vol.41 No.2
연구배경 : 종양괴사인자(tumor necrosis factor ; TNF)는 여러 암세포에 대해서 세포독성을 보임이 알려져 있다. 그리고, 최근 분자생물학적 발전에 힘입어 TNF 유전자를 암세포에 이입하고 발현시키는 연구가 그 동안 진행되었다. 이들 연구의 목적은, TNF를 전신적으로 쓸 경우 전신 부작용이 심하여 인체에 쓸 수 없는 현 단계에서 암세포 자체에서 TNF를 만들어 내어서 암세포 주위에서만 작용하게 함으로써 연체에 미치는 전신독성을 최소한으로 줄이고 암세포를 사멸시키는 것이었다. 암세포에 TNF 유전자를 이입함으로써 예상되는 항암효과가 성공을 거두기 위해서는 첫째, TNF 유전자가 이입된 암세포에서 분비된 TNF가 주위 암세포를 성공적으로 사멸시켜야 하고 둘째는 분비된 TNF가, TNF 유전자가 이입된 암세포 자신을 사멸시켜야 한다. 본 연구는 이 중 두번째 기전, 즉 TNF 유전자가 이입된 암세포가 자신이 분비한 TNF에 의해서 사멸되거나 또는 세포 독성이 나타날 수 있는가를 검증하는데 목적을 두었다. 방법 : TNF에 감수성을 보이는 인체의 중피종 세포주인 NCI-H2058과 생쥐 섬유육종 세포주인 WEHI164에 TNF-$\alpha$ 유전자를 retroviral vector를 이용하여 이입하고 TNF를 발현을 시도하였다. DNA 수준과 단백질 수준에서 TNF-$\alpha$ 유전자가 제대로 이입되어 발현되는지 PCR과 ELISA 및 bioassay(MTT assay)로 확인하였다. 그리고, TNF 유전자가 이입된 세포주가 자신이 분비하는 TNF에 사멸되는지 아니면 생존하는지 MTT(dimethylthiazolyl diphenyltetrazolium) assay로 알아보았다. 그리고, 만일 TNF 유전자가 이입된 암세포가 자신이 분비한 TNF에 사멸되지 않고 생존할 경우, TNF 유전자가 이입된 NCI-H2058-TNF와 WEHI164-TNF 암세포주가 외부에서 준 TNF에도 내성을 보이는지도 추가로 MTT assay 방법으로 확인해 보았다. 결과 : 1) TNF-$\alpha$ 유전자 이입 및 발현 확인 PCR을 시행한 결과, TNF 유전자가 이입된 NCI-H2058-TNF와 WEHI164-TNF 세포주는 790 base pair 크기의 진한 DNA band를 보인 반면 각각의 모세포주에서는 보이지 않아서 retroviral vector를 이용한 유전자 이입이 DNA 수준에서 이루어졌음을 확인할 수 있었다. 그리고, NCI-H2058-TNF와 WEHI164-TNF의 상층 배양액의 TNF양을 ELISA와 bioassay(MTT assay)로 측정한 결과, 생물학적 활성을 지닌 TNF를 각각 $23.6{\pm}0.84ng$/24h/$10^6cells$, $12.2{\pm}0.36ng$/24h/$10^6cells$ 생산함을 알 수 있었다. 2) TNF 유전자 이입 전후, 암세포의 TNF에 대한 감수성 비교 TNF 유전자 이입 전후의 TNF에 대한 세포주의 감수성(세포사망율)을 TNF의 농도 변화에 따라 비교한 결과, NCI-H2058의 경우 TNF 농도 100ng/ml에서 모세포는 $25{\pm}3%$의 세포독성을 보인 반면 TNF 유전자 이입 후에는 $3{\pm}2%$의 세포독성을 보여 통계적으로 유의한 차이가 있었다(p<0.01). 그리고, WEHI-164의 경우도 TNF 농도 100ng/ml에서 모세포는 $73{\pm}5%$의 세포독성을 보인 반면 TNF 유전자 이입 후에는 $3{\pm}2%$의 세포독성을 보여 통계적으로 유의한 차이가 있었다(p<0.01). 결론 : TNF에 감수성을 보이는 암세포주인 NCI-H2058과 WEHI164에 TNF 유전자 이입을 시행하고 TNF가 발현되게 하였을 때, TNF 유전자를 이입받은 두 암세포주 모두에서 자신이 생산해 내는 TNF에 내성을 보여 생존하였다. 뿐만 아니라 생존한 이들 세포는 외부에서 준 TNF에 대해서도 내성을 보였다. 따라서, 암세포에 대한 TNF 유전자 이입을 통한 유전자 요법이 성공 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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Developing a Diagnostic Bundle for Bronchiectasis in South Korea: A Modified Delphi Consensus Study
오연목,Hayoung Choi,Hyun Lee,나승원,Jong Geol Jang,Ji-Ho Lee,Byung Woo Jhun,Hye Yun Park,Ji Ye Jung,Seung Jun Lee,조경욱,Chin Kook Rhee,Changwhan Kim,이세원,Kyung Hoon Min,Yong-Soo Kwon,Deog Kyeom Kim,Jin Hwa Lee 대한결핵및호흡기학회 2022 Tuberculosis and Respiratory Diseases Vol.85 No.1
Background: Because the etiologies of bronchiectasis and related diseases vary significantly among different regionsand ethnicities, this study aimed to develop a diagnostic bundle for bronchiectasis in South Korea. Methods: A modified Delphi method was used to develop expert consensus statements on a diagnostic bundle forbronchiectasis in South Korea. Initial statements proposed by a core panel, based on international bronchiectasisguidelines, were discussed in an online meeting and two email surveys by a panel of experts (≥70% agreement). Results: The study involved 21 expert participants, and 30 statements regarding a diagnostic bundle for bronchiectasiswere classified as recommended, conditional, or not recommended. The consensus statements of the expert panel wereas follows: A standardized diagnostic bundle is useful in clinical practice; diagnostic tests for specific diseases, includingimmunodeficiency and allergic bronchopulmonary aspergillosis, are necessary when clinically suspected; initialdiagnostic tests, including sputum microbiology and spirometry, are essential in all patients with bronchiectasis, andpatients suspected with rare causes such as primary ciliary dyskinesia should be referred to specialized centers. Conclusion: Based on this Delphi survey, expert consensus statements were generated including specific diagnostic,laboratory, microbiological, and pulmonary function tests required to manage patients with bronchiectasis in SouthKorea.
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오연목 대한의사협회 2018 대한의사협회지 Vol.61 No.5
Asthma in the elderly tends to be underdiagnosed and undertreated despite its high morbidity and mortality. The diagnosis and management of asthma in the elderly is essentially the same as in adult patients. For diagnosis, spirometry is crucial, in addition to an assessment of typical symptoms; for management, the use of an inhaled corticosteroid is essential to control asthma. In elderly asthmatics, medications controlling asthma appear to be less efficacious than in younger asthmatics, and show more adverse effects. Other diseases with symptoms similar to asthma in the elderly are chronic obstructive pulmonary disease, tuberculous destroyed lung, bronchiectasis, lung cancer, and heart disease, and care should therefore be taken to differentiate these conditions from asthma. Vaccinations for influenza and pneumococcus are recommended for elderly individuals with asthma.
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오연목 대한의사협회 2014 대한의사협회지 Vol.57 No.2
Chronic obstructive pulmonary disease (COPD) is one of the highest ranking diseases with regard to prevalence andmortality in Korea and also worldwide. In the past decade, effective inhaler medications for COPD treatment have beendeveloped or approved. These inhaler medications have been proven to have beneficial effects on symptoms, lungfunction, quality of life, exercise capacity, and exacerbation. The inhalers used widely are long-acting anticholinergics,long-acting beta2-agonists, and combined inhalers of a corticosteroid and long-acting beta2-agonist. These inhalermedications are more effective than oral medications and less likely to produce adverse events. However, the inhalermedications should be used appropriately to achieve the desired effect. For COPD patients with a forced expiratoryvolume in 1 second (FEV1) less than 80% of the predicted value, a long-acting anticholinergic or long-acting beta2-agonist is usually the medication of first choice. If a COPD patient with a FEV1 less than 60% of the predicted valuesuffers frequent exacerbations, a combined inhaler of corticosteroid and long-acting beta2-agonist is a good choice. Toprescribe an inhaler medication for COPD patients, spirometry should be performed, not only to confirm the diagnosisbut also to define severity. These effective inhaler medications should be used widely for COPD patients in Korea.
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