Ophiopogonin D (OPD) is a steroidal glycoside derived from Ophiopogon japonicus , a traditional Chinese medicine with diverse biological activities, including antithrombosis, anti-inflammation, and antitussive effects. To investigate the cellular effects and mechanisms of OPD on oral squamous cell carcinoma, cell viability was explored, and the effects of OPD on cell cycle regulators, apoptotic marker proteins, and key proteins involved in metastasis and signaling pathways were examined by MTT assay and Western blotting in YD38 cells. OPD strongly inhibited cell proliferation and induced caspase-dependent apoptosis of YD38 cells by suppressing the cell cycle and activating caspase-3 and poly ADP ribose polymerase. Additionally, OPD suppressed the expression of vital proteins regulating metastasis and proliferation within the integrin/matrix metalloproteinases/FAK and AKT/PI3K/mTor pathways. Thus, OPD can be a potential treatment candidate for gingival cancer.

Introduction
Materials and Methods
    1. Chemicals and reagents
    2. Cell lines and cell culture
    3. MTT assay
    4. Western blot analysis
Results
    1. OPD selectively exerted cytotoxic effects againstYD38 cells
    2. OPD downregulated the cell cycle regulators
    3. OPD induced apoptosis on YD38 cells
    4. OPD suppressed metastasis through integrin/MMPs/FAK pathway
    5. OPD inhibited the PI3K/AKT/mTor and JNKpathways
Discussion
Funding
Conflicts of Interest
References

• Nguyen Thi Kieu Trang(Department of Pharmacology and Dental Therapeutics, College of Dentistry, Chosun University, Gwangju 61452, Republic of Korea, Department of Pharmacy, Thai Binh University of Medicine and Pharmacy, Thai Binh City 06000, Vietnam)
• Vu Phuong Dong(Department of Pharmacology and Dental Therapeutics, College of Dentistry, Chosun University, Gwangju 61452, Republic of Korea)
• Hoon Yoo(Department of Pharmacology and Dental Therapeutics, College of Dentistry, Chosun University, Gwangju 61452, Republic of Korea) Corresponding author