• Journal of Gynecologic Oncology
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• 제29권6호
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• pp.99.1-99.11
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• 2018

Objective: The present study is to evaluate the biological functions of long non-coding RNA (lncRNA), X-inactive specific transcript, X-inactive specific transcript (XIST) in human epithelial ovarian cancer (EOC). Methods: XIST was upregulated in EOC cell lines, CAOV3 and OVCAR3 cells by lentiviral transduction. The effects of XIST overexpression on cancer cell proliferation, invasion, chemosensitivity and in vivo tumor growth were investigated, respectively. Possible sponging interaction between XIST and human microRNA hsa-miR-214-3p was further evaluated. Furthermore, hsa-miR-214-3p was overexpressed in XIST-upregulated CAOV3 and OVCAR3 cells to evaluate its effect on XIST-mediated EOC regulation. Results: Lentivirus-mediated XIST upregulation had significant anticancer effects in CAOV3 and OVCAR3 cells by suppressing cancer cell proliferation, invasion, increasing cisplatin chemosensitivity and inhibiting in vivo tumor growth. Hsa-miR-214-3p was confirmed to directly bind XIST, and inversely downregulated in XIST-upregulated EOC cells. In EOC cells with XIST upregulation, secondary lentiviral transduction successfully upregulated hsa-miR-214-3p expression. Subsequently, hsa-miR-214-3p upregulation functionally reversed the anticancer effects of XIST-upregulation in EOC. Conclusion: Upregulation of lncRNA XIST may suppress EOC development, possibly through sponging effect to induce hsa-miR-214-3p downregulation

• Tuberculosis and Respiratory Diseases
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• 제42권3호
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• pp.302-313
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• 1995

연구배경: 종양괴사인자(Tumor necrosis factor; TNF)는 다양한 생물학적인 작용을 가지며 종양 세포에 대한 세포 독성은 그 대표적인 기능중의 하나이다. TNF-$\alpha$는 생체외에서(in vitro) 몇몇 종양 세포주에 대하여 항암제, 특히 topoisomerase II targeted chemotherapeutic agent의 세포 독성 효과를 상승적으로 증가시키는 것이 알려져 있다. 최근 암세포에 대한 cytokine 유전자 요법에서 TNF는 중요한 대상으로 여겨지고 있으며, 유전자 이입에 의해 암조직이 TNF를 생성하게 될 경우 암 증식 억제 효과가 있음이 보고되고 있다. 연구자는 암세포에 TNF-$\alpha$ 유전자를 이입하여 자신이 TNF-$\alpha$를 생성하도록 형질을 변환시킨 암세포는 topoisomerase II 억제 항암제에 대한 김수성에 변화가 있을 것이라는 가설을 수립하였고 이를 검증하고자 본 연구를 수행하였다. 본 연구에서는 생체외로(in vitro) TNF-$\alpha$ 유전자를 이입하여 TNF-$\alpha$를 생성하는 암세포주에서 topoisomerase II targeted drug에 대한 항암제 감수성 효과가 모세포주에 비하여 증대될 수 있는지를 알아 보고자하였다. 방법: TNF-$\alpha$에 감수성을 보이는 것으로 알려진 인체 중피종 세포주인 NCI-H2058 세포주 및 생쥐의 섬유육종 세포주인 WEHI164 세포주와 인체 비소세포 폐암 세포주인 A549 세포주를 배양하여, 먼저 임상에서 흔히 폐암의 항암 화학 요법 치료에 널리 쓰이는 대표적인 topoisomerase II targeted chemotherapeutic drug인 etoposide(VP-16)와 doxorubicin(adriamycin)을 가하였을 때 관찰된 세포 독성을 MTT assay로 측정하고, 각 모세포주(parenta1 cell line)에 TNF-$\alpha$의 유전자를 이입시켜서 형절 변환한 세포주(transformed cell line)에 대하여 각각 동일한 항암제를 가하였을 때 관찰된 세포 독성의 정도를 같은 방법으로 측정하여, 그 결과를 비교 분석하였다. 또한 모세포주에 외부에서 TNF를 가하여 전처치한 후 동일한 항암제를 가하였을 때의 세포독성을 관찰하여 비교 분석하였다. 결과: H2058 세포주에서는 TNF-$\alpha$ 유전자를 이입한 세포주 topoisomerase II targeted drug을 가하였을 때, 항암제 감수성이 모세포주에 같은 항암제를 가하였을 때에 비하여 의미있게 증가함을 관찰할 수 있었으나(p<0.05), WEHI 세포주와 A549 세포주에 있어서는 TNF-$\alpha$ 유전자를 이입한 세포주에서 모세포주에 비하여 항암제 감수성이 증가하지는 않았다. 결론: TNF-$\alpha$ 유전자의 이입이 topoisomerase II targeted chemotherapeutic drug에 대한 항암제 감수성을 증가시키는 효과는 세포주에 따라 다양한 결과를 보이는 것을 알 수 있었으며, 적어도 선택된 특정 종류의 호흡기계 암세포에 있어서는 TNF-$\alpha$ 유전자의 이입으로 항암제 감수성(chemosensitivity)을 증가시킬 수 있을 것으로 사료된다.

• 대한화장품학회지
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• 제15권1호
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• pp.37-50
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• 1989

The in Vitro chemosensitivity of fibroblast cell strains was determined using a semiautomated tetrazolium-based colorimetric assay(MTT assay) to 16 cosmetic materials. This assay is useful method to evaluate toxic effects of the chemicals. From assay results, we determined that the preservatives are more toxic than moisteurizers. The chemicals in the same group have a different toxicity. That is, in preservatives, Germall -115 is more toxic than Danisol -M, -p, and in surfactant sodium laurel sulfate than Myrj 52, and in moisteurizers, 1, 3-butylene glycol is more safe than the others. When the results from this assay for preservatives were compared with patch test results, good correlation was observed. Forthemore, this assay method can be used together with Patch test for the evaluation of the chemical toxicity, particularly in cosmetic field.

• Tuberculosis and Respiratory Diseases
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• 제62권1호
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• pp.33-42
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• 2007

연구배경: 다양한 고형암에서 HO-1의 높은 발현이 알려져 있고, 그것의 항산화와 항세포고사의 역할로 인해 빠른 암종의 성장에 중요한 역할이 있음이 보고되고 있다. 대표적인 활성산소종 생성 항암제인 Cisplatin은 현재까지 폐암치료에 가장 광범위하게 사용되고 있으나, 여러 내성발생이 임상치료의 주요문제로 대두되고 있다. 저자들은 A549 폐암세포주에서 HO-1의 발현이 증가되었고 HO-1 활성억제제나 siRNA 방법을 통해 생존율의 의미 있는 감소와 세포고사가 유도됨을 보고한 바 있다. 이 연구의 목적은 A549 폐암세포주에 cisplatin 처리시 HO-1의 발현의 증가유무와 기전을 규명하고 실제 HO-1의 억제가 cisplatin에 의한 항암제 감수성을 증가시키는지를 알아보는데 있다. 방 법: 비소세포폐암세포주인 A549, NCI-H23, NCIH157, NCI-H460을 이용하였다. 세포독성은 MTT 방법으로 구하였고, HO-1, Nrf2, MAPK의 발현은 Western blotting으로 확인하였다. 또한 MAPK억제제들을 전처치한 후 cisplatin에 의해 유도된 Nrf2와 HO-1의 발현에 미치는 영향을 역시 Western blotting으로 관찰하였다. A549세포에 활성억제제인 ZnPP나 HO-1 small interfering RNA (siRNA)을 주입하여 cisplatin과의 병합요법시 생존율의 배가효과 유무를 MTT 방법으로 확인하였고, 이러한 효과가 ROS 형성과 HO-1의 발현변화와 관련되는지를 알아보기 위해 $carboxy-H_2DCFDA$ 방법과 Western blotting을 통해 각각 확인하였다. 결 과: Cisplatin 처리시 다른 세포주에 비해 A549 세포가 의의 있게 내성을 보였다. $10{\mu}M$의 농도에서 시간 의존적으로 HO-1, Nrf2, MAPK의 발현이 증가하였고, MAPK 억제제들을 전 처치하였을 때 cisplatin에 의해 유도된 HO-1과 Nrf2의 발현이 억제됨을 확인하였다. HO-1의 활성억제제인 ZnPP와 HO-1 siRNA를 통해 HO-1 mRNA를 직접 억제하는 방법으로 cisplatin과 병합치료시 단독치료에 비해 의의 있는 생존율의 감소를 보였다. 이러한 효과는 활성산소종의 생성 증가와 HO-1의 발현억제에 의한 결과임을 확인하였다. 결 론: Cisplatin 처리시 HO-1의 발현은 MAPKNrf2-HO-1의 경로를 통해 증가하였고, 부분적으로 치료에 대한 내성과 관련이 있었으며, ZnPP 등의 활성억제제나 siRNA를 통한 knock-down 방법으로 HO-1 을 표적으로 억제하는 치료방법을 통해 cisplatin의 항암제 감수성을 증가시켰다.

• Archives of Pharmacal Research
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• 제20권5호
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• pp.410-413
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• 1997

In the course of screening synthetic compounds to inhibit tumor cell growth, pyrrolo[1,2-.alpha.] benzimidazole (PBI), an intermediate of azamitosene, was found to inhibit a proliferation of gastric cancer cell lines. Despite a potential cytotoxic activity against solid tumor cells as opposed to that against rapidly-doubled leukemic cells, there has been no report on the inhibition of gastric cancer cell line by PBI and its' derivatives. The present experiment was designed to determine if PBI derivatives can effectively inhibit the cellular proliferation of gastric cancer cells by using in vitro as well as in vivo chemosensitivity system (MTT assay, clonogenic assay and human tumor xenografted assay). Of the tested PBI derivatives, PBI (18) and PBI (20), displayed the effective growth inhibition of cultured gastric cancer cells or even in the xenografted nude mouse model.

• Asian Pacific Journal of Cancer Prevention
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• 제13권2호
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• pp.689-692
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• 2012

Objective: To investigate correlations between adenosine triphosphate chemotherapy response assay (ATP-CRA) and clinical outcomes after ATP-CRA-based chemotherapy for drug selection in patients receiving intravesical chemotherapy to prevent recurrence of superficial bladder cancer after surgery. Methods: The chemosensitivities of 12 anticancer drugs were evaluated, including 5-Fu ADM, and EPI, using ATP-CRA and primary tumor cell culture in 54 patients. In addition, a further 58 patients were treated according to clinical experience. Differences in post-chemotherapeutical effects between drug sensitivity assay and experience groups were compared. Results: The evaluable rate of the test was 96.3%, the clinical effective rate was 80.8%, the sensitivity rate was 97.6% (41/42), the specificity was 20%, the total predicting accuracy was 74.3%, the positive predictive value was 83.7% (41/49), the negative predictive value was 66.7% (2/3); in the drug sensitivity test group, the clinical effective rate was 80.8%, the experience group response rate was 63.8%, with a significant difference in clinical effects between the ATP-based sensitivity and experience groups (${\chi}^2$=7.0153, P<0.01). Conclusion: ATP-CRA is a stable, accurate and potentially practical chemosensitivity test providing a predictor of chemotherapeutic response in patients with superficial bladder cancer.

• Asian Pacific Journal of Cancer Prevention
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• 제14권12호
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• pp.7375-7379
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• 2013

The main aim of this study was to investigate the roles of GST-${\pi}$ and $pol{\beta}$ genes in the chemoresistance of esophageal carcinoma cells. Eukaryotic expression vectors containing each gene were constructed and transfected into EC9706 cells, and the biological effects of the two genes assessed based on a resistance index. We additionally investigated the in vitro and in vivo anti-resistance effects of GST-${\pi}$ and $pol{\beta}$ genes using recombinant lentiviruses carrying siRNAs against the two genes. Our results showed that upregulation of GST-${\pi}$ and $pol{\beta}$ genes suppresses chemosensitivity of esophageal carcinoma cells to cisplatin, while downregulation of these two genes with RNAi technology reverses this chemoresistance. Multi-site injection of recombinant lentivirus targeting the GST-${\pi}$ gene into transplanted cDDP tumors effectively reversed their chemoresistant phenotype. However, the same treatment against the $pol{\beta}$ gene did not lead to significant efficacy against chemoresistance.

• Journal of Microbiology and Biotechnology
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• 제14권2호
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• pp.350-355
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• 2004

Sulforhodamine B (SRB) assay is a rapid, sensitive, and inexpensive method for measuring cell proliferation and chemosensitivity. However, the lactate dehydrogenase (LDH) release assay is generally used to measure cytototoxicity of infectious microorganisms against host cells. In this study, we investigated the possibility of applying the SRB assay to determine cytotoxicity for infectious microorganisms, and compared the results with those obtained by the LDH release assay. We used Vibrio vulnificus as a model of infectious microorganisms. The SRB assay showed that V vulnificus strongly induced cytotoxic activity against human intestinal cells, Caco-2 and INT-407 cells. The degree of cytotoxicity closely correlated with infection time and number ratios of V. vulnificus to intestinal cells (MOI, multiplicity of infection). Furthermore, cytotoxicity values obtained by SRB assay correlated well with results obtained by the LDH release assay, and both assays gave a linear response with respect to MOI Heat-inactivation of V. vulnificus for 35 min at $60^{\circ}C$ did not induce cytotoxic activity, indicating that viability of V. vulnificus is crucial for cytotoxic activity against intestinal cells. Although both assays are suitable as cytotoxicity endpoints, the SRB assay is recommended for measuring cytotoxicity of infectious microorganisms against host cells because of its significantly lower cost and more stable endpoint than the LDH release assay.

• 치과방사선
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• 제25권2호
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• pp.331-341
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• 1995

The purpose of this study was to aid in the prediction of tumor cell tolerance to radiotherapy and/or chemotherapy. For this study, cell surviving curves were obtained for murine melanoma Bl6 cell line using semiautomated M1T assay. 2,4,6,8, 10Gy were irradiated at a dose rate of 210cGy/min using /sup 60/Co Irradiator ALOORADO 8. After irradiatior, B16 cell lines(2.5×10⁴ cells/ml) were exposed to bleomycin and cisplatin at concentration of 0.2㎍/㎖, 2㎍/㎖ and 20㎍/㎖ for I hour respectively. The viable cells were determined for each radiation dose and/or each concentration of drug. And they were compared to control values. The obtained results were as follows : 1. There was significant difference of surviving fraction at 4, 6, 8, 10Gy on B16 cell line(P<0.05). 2. There was significant difference of cytotoxicity between bleomycin and cisplatin at concentration of 0.2㎍/㎖ and 2㎍/㎖(P<0.05) on B16 cell line, but there was no significant difference of cytotoxicity at concentration of 20㎍/㎖ on B16 cell line. 3. There was significant difference of cytotoxicity of bleomycin after irradiation of 2Gy and 10Gy on B16 cell line(P<0.01). 4. There was significant difference of cytotoxicity of cisplatin at concentration of 20㎍/㎖ after irradiation on B16 cell line.

• Journal of Microbiology and Biotechnology
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• 제26권8호
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• pp.1490-1503
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• 2016

Colorectal cancer (CRC) is the third most common cancer in the world. Although 5-fluorouracil (5-FU) is the representative chemotherapy drug for colorectal cancer, it has therapeutic limits due to its chemoresistant characteristics. Colorectal cancer cells can develop into cancer stem cells (CSCs) with self-renewal potential, thereby causing malignant tumors. The human gastrointestinal tract contains a complex gut microbiota that is essential for the host's homeostasis. Recently, many studies have reported correlations between gut flora and the onset, progression, and treatment of CRC. The present study confirms that the most representative symbiotic bacteria in humans, Lactobacillus plantarum (LP) supernatant (SN), selectively inhibit the characteristics of 5-FU-resistant colorectal cancer cells (HT-29 and HCT-116). LP SN inhibited the expression of the specific markers CD44, 133, 166, and ALDH1 of CSCs. The combination therapy of LP SN and 5-FU inhibited the survival of CRCs and led to cell death by inducing caspase-3 activity. The combination therapy of LP SN and 5-FU induced an anticancer mechanism by inactivating the Wnt/β-catenin signaling of chemoresistant CRC cells, and reducing the formation and size of colonospheres. In conclusion, our results show that LP SN can enhance the therapeutic effect of 5-FU for colon cancer, and reduce colorectal cancer stem-like cells by reversing the development of resistance to anticancer drugs. This implies that probiotic substances may be useful therapeutic alternatives as biotherapeutics for chemoresistant CRC.