SiHa 자궁경부암 세포주에서 인유두종바이러스 16형 E6/E7 siRNA에 의한 세포성장 억제 비교
Comparison of cell growth suppression in SiHa cervical carcinoma cell line by human papillomavirus type 16 E6/E7 siRNAs
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목적: 인유두종바이러스 (Human papillomavirus, HPV)는 자궁경부암과 밀접한 관계가 있는 것으로 알려져 있으며, 특히 인유두종바이러스 16형은 고위험군 바이러스로 분류된다. 인유두종바이러스 16형의 암 유전자 E6와 E7의 발현 저해에 따른 자궁경부암 세포주내의 발암 유전자 특이적 변화를 확인하고자 하였다. 연구 방법: 인유두종바이러스 16형을 발현하는 자궁경부암 SiHa 세포주에서 E6 유전자에 특이적으로 작용하는 인유두종바이러스 16형 E6 siRNA (siRNA 377)와 E6와 E7 유전자에 동시에 작용하는 인유두종바이러스 16형 E6 siRNA (siRNA 3)와 인유두종바이러스 16형 E7 siRNA (siRNA 198)를 사용하였다. 각각의 siRNA를 SiHa 세포주에 이입 (transfection)시킨 후, 24시간과 48시간 후에 세포수 및 세포형태의 변화를 관찰하였다. 또한 E6와 E7에 대한 RT-PCR을 실시하여 인유두종바이러스 16형 E6, E7 siRNA의 유전자 발현 저해 정도를 확인하였다. 결과: 3종류의 siRNA 모두 세포성장 저해 효과를 나타내었으며, siRNA 377에 비해 siRNA 3와 siRNA 198의 세포성장 억제 효과가 더 크게 나타났다. siRNA 377은 E6 mRNA만의 발현을 감소시켰고 siRNA 3과 siRNA 198은 E6와 E7 mRNA의 발현을 동시에 감소시켰다. 결론: 인유두종바이러스 16형이 감염된 자궁경부암 SiHa 세포주에서 E6 유전자만의 억제보다는 E6와 E7 유전자를 동시에 억제하는 것이 더 큰 세포 성장 억제를 보였다.
Abstract
Objective: Human cervical cancer is caused by the high-risk types of human papillomavirus (HPV) such as HPV16, which possess the E6 and E7 oncogenes, whose expressions are a prerequisite for cancer development. We performed this study to compare the efficacy of antitumor activity by HPV siRNA which silences only E6 or both E6/E7. Methods: We transfected siRNA 377 (HPV16 E6 siRNA), siRNA 3 (HPV16 E6 siRNA), and siRNA 198 (HPV16 E7 siRNA) into SiHa cell line (siRNA 377 silences only E6, and siRNA 3 and siRNA 198 silence both E6 and E7). We experimented cell counts and morphologic changes 24 and 48 hours after transfection and expressions of HPV16 E6/E7 mRNA by RT-PCR. Results: siRNA 377, siRNA 3, and siRNA 198 suppressed the cell growth. siRNA 3 and siRNA 198 were more potent than siRNA 377 in cell growth suppression. siRNA 377 knocked down the expression of E6 mRNA, and both siRNA 3 and siRNA 198 knocked down the expression of E6/E7 mRNA. Conclusion: Our results suggest that simultaneous suppression of E6 and E7 was more potent than E6-specific suppression in cancer cell growth.
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