High Pressure Processing Assisted Enzymatic Hydrolysis of Corbicula fluminea and Evaluation of Its Anti-cancer Activity
學生姓名:
陳佳琳
指導教授:
張君如
學期:
110下
摘 要:
Corbicula fluminea hydrolysate shows physiology functions such as hepatoprotection, antioxidant, and anticancer. High pressure processing (HPP) can improve the degree of hydrolysis and increase the biological activity of peptides. This study aims to use HPP-assisted enzymatic hydrolysis of C. fluminea (pCFH), and to evaluate pCFH effects on anticancer ability in human triple negative breast cancer (TNBC) MDA-MB-231 cells. Response surface methodology found that the optimum condition for pCFH preparation was 40 grams of C. fluminea powder hydrolyzed with 2% of Umamizyme G for 3 hours, 1% of pepsin for 1 hour, and extracted under 450 MPa for 15 minutes. The contents of soluble protein and peptide were 1.98 mg/mL and 450.98 mg/mL, respectively; and the IC50 of pCFH on MDA-MB-231 cells was 0.89 mg/mL. Subsequently, treatment with doxorubicin (IC50 215 nM) and supplied with pCFH (0.45, 0.90, and 1.80 mg/mL) inhibited the cell viability to 86.2%, 73.1%, and 62.8% of control. Treatment with paclitaxel (IC50 45.6 nM) and supplied with pCFH (0.45, 0.90, and 1.80 mg/mL) also decreased the cell viability to 85.8%, 70.8%, and 58.3% of control. Annexin V and PI double staining demonstrated that treatment with pCFH (0.45, 0.90, and 1.80 mg/mL) for 36 hours increased early apoptotic cells to 188.4%, 224.0%, and 259.8% of control. Furthermore, treatment with pCFH (0.45, 0.90, and 1.80 mg/mL) reduced wound healing (90.2%, 55.3%, and 49.3% of control), and cell migration (67.8%, 60.4%, and 40.4% of control). In summary, pCFH increased chemosensitivity, and inhibited cell migration, as well as induced cell apoptosis of TNBC cells. Hence, pCFH is a promising dietary agent for attenuation of TNBC.
摘要.pdf