[논문]HeLa S3 자궁암 세포에서 paclitaxel 에 의해 유도된 Poly(ADP-ribose) Polymerase 분철과 세포자멸사와의 관계

장정현; 김광연; 안순철; 권헌영

doi:10.5352/jls.2007.17.8.1027

HeLa S3 자궁암 세포에서 paclitaxel 에 의해 유도된 Poly(ADP-ribose) Polymerase 분철과 세포자멸사와의 관계
Relation of Poly(ADP-ribose) Polymerase Cleavage and Apoptosis Induced by Paclitaxel in HeLa S3 Uterine Cancer Cells 원문보기

생명과학회지 = Journal of life science, v.17 no.8 = no.88, 2007년, pp.1027 - 1033

장정현 (부산카톨릭대학교 보건과학대학 임상병리학과) , 김광연 (부산대학교 미생물학 및 면역학교실) , 안순철 (부산대학교 미생물학 및 면역학교실) , 권헌영 (부산카톨릭대학교 보건과학대학 임상병리학과)

초록
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Paclitaxel이 암세포에서 세포예정사를 유발할지라도, 아직 정확한 기전은 잘 알려져 있지 않다. 이에 본 연구에서는 HeLa $S_{3}$ 자궁암세포에서의 paclitaxel이 어떠한 영향을 미치는지 알아보고자 한다. 그리하여 방법으로는 세포독성검사, apoptotic cells의 형태학적 변화(DAPI 염색 ), western blot 분석법을 사용하여 수행하였다. 본 연구의 결과로 paclitaxel은 HeLa $S_{3}$ 세포에서 세포독성을 보이며 특히 paclitaxel의 $IC_{50}$ 값은 약 1 ${\mu}M$이며, paclitaxel 처리한 HeLa $S_{3}$ 세포에서 형태학적 변화(분절화)를 관찰하였고, flow cytometric 분석에서는 G2/M기가 차단되어 paclitaxel은 세포주기 특히 Sub-$G_{1}$기를 조절함을 알 수 있다. 그리고 Paclitaxel을 처리한 HeLa $S_{3}$ 세포에서는 PARP cleavage를 유발하였고 Bc1-2의 감소와도 관련되었다.

Abstract ▼ AI-Helper

Although paclitaxel induces apoptosis of cancer cells, its exact mechanism of action is not yet known. The present study has been performed to determine whether influence of paclitaxel in HeLa $S_{3}$ uterine cancer cells. Three assays were employed in this study: cell cytotoxicity, morphological assessments of apoptotic cells (DAPI staining assay), and western blot analysis. The results indicated that paclitaxel has cytotoxic effects in HeLa $S_{3}$ cells. Especially, the $IC_{50}$ value of paclitaxel was about 1 ${\mu}M$. And morphological changes (fragmentation) of cells were observed by paclitaxel in HeLa $S_{3}$ cells. The flow cytometric analysis of paclitaxel-treated cells indicated a block of G2/M phase. The results that pacli-taxel regulates the cell cycle, especially Sub-$G_{1}$ phase. Paclitaxel induces apoptosis of HeLa $S_{3}$ cells via PARP-dependent fashion, and this apoptosis is related to disappearance of Bcl-2 proteins.

주제어

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문제 정의

These results also support this notion. This study is the first to provide experimental evidence demonstrating that a dose-dependent cytotoxic effect by paclitaxel in HeLa S3 cells. Also, cell viability of the other cancer cells decreased in a dose-dependent by paclitaxel that was observed.

제안 방법

Although much has been learned concerning the metabolism of paclitaxel, a clear understanding of the biochemical bases for its apoptosis-inducing activity have not yet emerged. The purpose of this study is to elucidate the fundamental mechanism in relation to apoptosis induced by paclitaxel in HeLa S3 cells.

이론/모형

0), 1% Triton X-100, 1 mM PMSF, 20 mg/mL aprotinin, 50 μg/mL leupeptin, 1 mM benzaidine, 1 mg/mL pep- statin) for 30 min, followed centrifugation (12,000 rpmz 30 min). Protein content was determined using the bradford assay. Equivalent amounts of protein (30 pg) were resolved by 12% SDS-PAGE, transferred to poly-vinylidene difluoride membrane, and probed with the antibodies.

성능/효과

Taken together, these results lead to the conclusion that the effect of paclitaxel in inducing apoptosis in HeLa S3 cells that is dependent cell cycle arrest and PARP activation. In conclusion, all observations indicate that paclitaxel may provide a superior therapeutic index and advantage in the clinic for the treatment of cervical cancer.
Taken together, these results lead to the conclusion that the effect of paclitaxel in inducing apoptosis in HeLa S3 cells that is dependent cell cycle arrest and PARP activation. In conclusion, all observations indicate that paclitaxel may provide a superior therapeutic index and advantage in the clinic for the treatment of cervical cancer.

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