Obstetrics & gynecology science

Korean Society of Obstetrics and Gynecology (대한산부인과학회)
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Apigenin-induced apoptosis in cervical cancer cell lines

자궁경부암 세포주에서 Apigenin에 의한 세포사 유도

Oh, Eun-Kyung;Kim, Hyun-Jung;Bae, Su-MI;Park, Mi-Young;Kim, Yong-Wook;Kim, Tae-Eung;Ahn, Woong-Shick
오은경;김현정;배수미;박미영;김용욱;김태응;안웅식

  • Published : 2008.08.15
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Abstract

Objective: Apigenin is a widely distributed plant flavonoid and was proposed as a potent antitumor agent. In this study, we investigated the anticancer effects of apigenin on human cervical cancer cell lines. For this, the effects of apigenin on growth inhibition and apoptosis were examined and also mRNA expression of E6 and E7, the main causes of development of cervical cancer, was also evaluated. Methods: To observe the anti-proliferative effects, cervical cancer cell lines, 5×103 cells/well (96 well plate) of Caski, HeLa and C33A were plated and 24 h later treated with apigenin for three days and then MTT assay was performed. For apoptosis analysis, Annexin V-FITC staining was performed. To examine the effect on anchorage-independent growth by apigenin, soft agar assay was performed. The mRNA expression of HPV E6/E7 was examined by quantitative RT-PCR. Results: Apigenin inhibited the growth of all three kind of cervical cancer cell lines (CaSki, HeLa, and C33A) and induced apoptosis in these cell lines. Also, anchorage-independent growth of Caski cells in soft agar was inhibited approximately 3 folds by apigenin treatment. Unexpectedly, mRNA expression level of both E6 and E7 in HeLa cells was not significantly affected by apigenin. Conclusion: These studies showed that apigenin inhibits cervical cancer cell growth through the induction of apoptosis. However, mRNA expression of HPV E6/E7 genes was not affected by the treatment of apigenin, indicating that the anti-cancer effect of apigenin in cervical cancer might be mediated via other pathway. Taken together, these results suggest that apigenin may provide a new therapeutic approach for cervical cancer.

목적: Apigenin은 보편적인 식물 방향족 화합물로서 항종양 효과를 가지는 것으로 알려져 있다. 본 연구는 자궁경부암 세포주 에서 apigenin의 세포성장 억제 효과 및 세포사멸 효과를 조사하고, HPV E6/E7 유전자의 발현에 미치는 영향을 조사하고자 하였다. 방법: 자궁경부암 세포주들에서 apigenin에 의한 항암효과를 확인하기 위하여 세포성장 억제 및 세포사멸 효과를 조사하고, HPV E6/E7의 발현을 조사하였다. 5×103 cells/well (96 well plate)로 CaSki, HeLa 그리고 C33A 세포를 분주한 후 24시간 뒤 apigenin을 농도별로 처리한 후 3일 동안 세포성장 억제효과를 MTT 측정법으로 확인하였으며, 세포사는 Annexin V-FITC staining을 실시하여 확인하였다. Caski 세포의 Anchorage-independent growth에 apigenin이 미치는 효과를 보기 위해 soft agar assay를 수행하였다. 또한 CaSki, HeLa 그리고 C33A 세포주에서 qRT-PCR을 통해 HPV E6/E7 유전자의 발현변화를 확인하 였다. 결과: Apigenin은 Caski, HeLa, C33A 세종류의 자궁경부암 세포주의 농도의존적으로 세포성장을 억제하고 세포 사멸을 유도하 였으며 또한 soft agar에서 Caski 세포의 anchorage-independent growth를 약 3배 정도 저해하였다. 예상과는 달리 Apigenin은 자궁경부암 세포주에 감염된 인유두종바이러스의 암유전자인 E6와 E7의 mRNA level에는 영향을 주지 않았다. 결론: CaSki, HeLa 그리고 C33A와 같은 자궁경부암 세포주에서 apigenin 투여는 암세포의 성장을 저해하고, 세포사를 유도함을 확인하였다. apigenin은 E6와 E7의 mRNA 발현에는 영향을 미치지 않았다. 따라서 자궁경부암 세포주에서의 apigenin의 항암 기전은 E6와 E7의 전사 후 또는 단백질 수준 또는 다른 발암 신호전달체계를 저해함으로써 항암효과를 나타내는 것으로 생각된다.

Keywords

References

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