Fangchinoline Exerts Anticancer Effects on Colorectal Cancer Cells by Evoking Cell Apoptosis via Endoplasmic Reticulum Stress

by Selleckchem | Jul 04 2023

We studied the anti-tumor effect of fangchinoline (FAN) against human colorectal cancer cell lines CCL-244 and SW480 and analyzed the mechanism of FAN action. The cell viability and apoptosis were assessed by MTT test and Annexin V-PI staining; caspase-3 activity was measured by Western blotting. The expression of endoplasmic reticulum stress-related proteins was assessed by real-time PCR, Western blotting, and gene transfection. It was found that FAN inhibited cell growth and induced apoptosis in human colorectal cancer cell lines CCL-244 and SW480 in a dose-dependent manner. The caspase-3 inhibitor Ac-DEVD-CHO could reverse the inhibitory effect of FAN. Moreover, FAN significantly increased the expression of endoplasmic reticulum stress-related proteins p-PERK, p-eIF2α, ATF4, and CHOP in CCL-244 and SW480 cells. In addition, endoplasmic reticulum stress inhibitor 4-phenylbutyric acid or CHOP knockdown could prevent FAN-induced apoptosis. Thus, FAN induced apoptosis of human colorectal cancer through activation of endoplasmic reticulum stress.

Comments:

The study you described investigated the anti-tumor effect of fangchinoline (FAN) on human colorectal cancer cell lines CCL-244 and SW480, and it aimed to elucidate the mechanism underlying FAN's action. Here's a summary of the findings:

FAN inhibited cell growth and induced apoptosis: The researchers observed that FAN treatment led to a dose-dependent inhibition of cell growth in both CCL-244 and SW480 colorectal cancer cell lines. Additionally, FAN induced apoptosis, a programmed cell death process, in these cell lines.

Caspase-3 involvement: Caspase-3 is a crucial enzyme involved in apoptosis. The researchers measured caspase-3 activity using Western blotting and found that FAN treatment increased caspase-3 activity. To further confirm the role of caspase-3, they used a caspase-3 inhibitor called Ac-DEVD-CHO, which was able to reverse the inhibitory effect of FAN, suggesting that caspase-3 activation is involved in FAN-induced apoptosis.

Activation of endoplasmic reticulum (ER) stress: The researchers explored the mechanism by which FAN induced apoptosis and focused on endoplasmic reticulum stress. They assessed the expression of ER stress-related proteins using real-time PCR, Western blotting, and gene transfection techniques. FAN treatment significantly increased the expression of several ER stress-related proteins, including p-PERK, p-eIF2α, ATF4, and CHOP, in both CCL-244 and SW480 cells.

Role of ER stress in FAN-induced apoptosis: To further investigate the involvement of ER stress in FAN-induced apoptosis, the researchers utilized an ER stress inhibitor called 4-phenylbutyric acid and performed CHOP knockdown experiments. They found that treatment with either the ER stress inhibitor or CHOP knockdown prevented FAN-induced apoptosis, suggesting that FAN induces apoptosis in colorectal cancer cells through the activation of ER stress, specifically involving the CHOP protein.

In summary, the study demonstrated that fangchinoline (FAN) inhibits cell growth and induces apoptosis in human colorectal cancer cell lines. The mechanism underlying FAN's action involves the activation of endoplasmic reticulum stress, as evidenced by the upregulation of ER stress-related proteins and the prevention of FAN-induced apoptosis by ER stress inhibition or CHOP knockdown. These findings provide insights into the potential therapeutic application of FAN for colorectal cancer treatment.