Humulus japonicus (HJ) is a widely used herbal medicine for pulmonary tuberculosis, hypertension, leprosy, and venomous wounds in Asia, particularly in China. Although HJ has certain physiological activities, such as longitudinal bone growth, antioxidation and alleviation of rheumatism, its anticancer activities, other than in colorectal and ovarian cancer, are yet to be studied. In this study, we investigated the anti-cancer activity and mechanism of methanol extracts of HJ (MeHJ) against human FaDu hypopharyngeal squamous carcinoma cells. MeHJ suppressed FaDu cell viability without affecting normal cells (L929), which was demonstrated using the MTT and Live & Dead assays. Furthermore, MeHJ effectively inhibited colony formation of FaDu cells, even at non-cytotoxic concentrations, and significantly induced apoptosis through the proteolytic cleavage of caspase-9, -3, -7, poly (ADP-ribose) polymerase and through the downregulation of BCL-2 and upregulation of BAX in FaDu cells, as determined by DAPI staining, flow cytometry, and western blot analyses. Collectively, these findings suggest that the inhibitory effects of MeHJ on the growth and colony formation of oral cancer cells may be mediated by caspase- and mitochondrial-dependent apoptotic pathways in human FaDu hypopharyngeal squamous carcinoma cells. Therefore, MeHJ has the potential to be used as a natural chemotherapeutic drug against human oral cancer.

Introduction
Materials ans Methods
    1. 실험재료
    2. 환삼덩굴 메탄올추출물 제조(MeHJ)
    3. 세포 배양
    4. 세포 성장률 분석
    5. Colony formation 측정
    6. Live & Dead assay
    7. DAPI 염색
    8. 유세포 분석
    9. 단백 발현 분석
    10. 통계적 유의성
Results
    1. MeHJ에 의해 FaDu 암세포의 성장 억제 및 세포사멸 유도
    2. 유세포 분석을 이용한 MeHJ에 의한 FaDu의 세포사멸
    3. MeHJ에 의한 세포사멸 관련 단백질 발현 유도
Discussion
Acknowledgements
Conflicts of Interest
References

• Ji Yeon Jang(Department of Oral Biochemistry, College of Dentistry, Chosun University, Gwangju 61452, Republic of Korea)
• Bo-Ram Park(Department of Dental Hygiene, College of Health and Welfare, Kyungwoon University, Gumi 39160, Republic of Korea)
• Seul Ah Lee(Department of Oral Biochemistry, College of Dentistry, Chosun University, Gwangju 61452, Republic of Korea)
• Mi Suk Choi(Department of Dental Hygiene, Chodang University, Muan 58530, Republic of Korea)
• Chun Sung Kim(Department of Oral Biochemistry, College of Dentistry, Chosun University, Gwangju 61452, Republic of Korea) Corresponding author