Though Farnesiferol C (FC) derived from Ferula asafoetida is known to have antiangiogenic and apoptotic effect in gastric, breast, nonsmall lung cancers, the underlying antitumor mechanism of FC is not fully understood so far. Hence, in the current study, apoptotic mechanism of FC was explored in colon cancers in association with carbon catabolite repression 4-negative on TATA-less 2 (CNOT2)/c-Myc signaling. Herein FC significantly increased cytotoxicity and reduced the number of colonies in HCT116 cells more effectively than in SW480 cells, though FC enhanced sub-G1 cell population in HCT116 and SW480 cells compared to untreated control. Consistently, FC activated the cleavages of Poly ADP-ribose polymerase (PARP) and Bax and attenuated the expression of pro-PARP and Cyclin D1 in HCT116 cells better than SW480 cells. Also, FC significantly reduced the expression of CNOT2 and c-Myc. Also, FC reduced of c-Myc stability in HCT116 cells by cycloheximide assay. Notably, CNOT2 depletion reduced the expression of c-Myc, while c-Myc depletion also attenuated the expression of CNOT2 in HCT116 cells, implying the crosstalk between CNOT2 and c-Myc. Furthermore, overexpression of c-Myc or CNOT2 promoted the expression of pro-PARP in HCT116 cells. Overall, these findings suggest that FC induces apoptosis via inhibition of CNOT2 and c-Myc in colon cancers for a potent anticancer candidate for further agriculture cultivation in Korea.

Abstract
Introduction
Materials and methods
    1. Preparation of Farnesiferol C (FC)
    2. Cell culture
    3. Cytotoxicity assay
    4. Colony formation assay
    5. Cell cycle analysis
    6. Western blotting
    7. RNA interference
    8. Cycloheximide assay
    9. Statistical analysis
Results and Discussion
    1. Cytotoxic and antiproliferative effect of FC inhuman colon cancer cells
    2. Effect of FC on sub G1 population and apoptosisrelated proteins in human colon cancer cells
    3. FC effectively regulated apoptosis related proteins anddiminished the stability of c-Myc in colon cancer cells
    4. The crosstalk between CNOT2 and c-Myc incolon cancer cells
    5. Overexpression of c-Myc or CNOT2 disturbs theapoptotic effect of FC to reduce pro-PARP in HCT116cells
Acknowledgments
References

• Deok Yong Sim(Research Professor, College of Korean Medicine, Kyung Hee University, Seoul 02447, Korea)
• Ji Sung Hwang(Doctoral Student, College of Korean Medicine, Kyung Hee University, Seoul 02447, Korea)
• Su Yeon Park(Doctoral Student, College of Korean Medicine, Kyung Hee University, Seoul 02447, Korea)
• Bum Sang Shim(Professor, College of Korean Medicine, Kyung Hee University, Seoul 02447, Korea)
• Sung Hoon Kim(Professor, College of Korean Medicine, Kyung Hee University, Seoul 02447, Korea)
• Jin Suk Koo(Professor, Department of Forest Science, Andong National University, Andong 36729, Korea) Corresponding author