Carvacrol instigates intrinsic and extrinsic apoptosis with abrogation of cell cycle progression in cervical cancer cells: Inhibition of Hedgehog/GLI signaling cascade

Afza Ahmad; Rohit Kumar Tiwari; Mohd Saeed; Hadba Al-Amrah; Ihn Han; Eun-Ha Choi; Dharmendra K Yadav; Irfan Ahmad Ansari

doi:10.3389/fchem.2022.1064191

Carvacrol instigates intrinsic and extrinsic apoptosis with abrogation of cell cycle progression in cervical cancer cells: Inhibition of Hedgehog/GLI signaling cascade

Front Chem. 2023 Jan 11:10:1064191. doi: 10.3389/fchem.2022.1064191. eCollection 2022.

Authors

Afza Ahmad¹, Rohit Kumar Tiwari¹, Mohd Saeed², Hadba Al-Amrah³, Ihn Han⁴, Eun-Ha Choi⁴, Dharmendra K Yadav⁵, Irfan Ahmad Ansari¹

Affiliations

¹ Department of Biosciences, Integral University, Lucknow, India.
² Department of Biology, College of Sciences, University of Hail, Hail, Saudi Arabia.
³ Department of Biological Sciences, Faculty of Science, King Abdulaziz University, Jeddah, Saudi Arabia.
⁴ Plasma Bioscience Research Center, Applied Plasma Medicine Center, Department of Electrical & Biological Physics, Kwangwoon University, Seoul, South Korea.
⁵ Department of Pharmacy and Gachon Institute of Pharmaceutical Science, College of Pharmacy, Gachon University, Incheon, South Korea.

Abstract

Recent times have seen a strong surge in therapeutically targeting the hedgehog (HH)/GLI signaling pathway in cervical cancer. HH signaling pathway is reported to be a crucial modulator of carcinogenesis in cervical cancer and is also associated with recurrence and development of chemoresistance. Moreover, our previous reports have established that carvacrol (CAR) inhibited the proliferation of prostate cancer cells via inhibiting the Notch signaling pathway and thus, it was rational to explore its antiproliferative effects in cervical cancer cell lines. Herein, the present study aimed to investigate the anticancer and apoptotic potential of CAR on C33A cervical cancer cells and further explore the underlying mechanisms. We found that CAR significantly suppressed the growth of C33A cells, induced cell cycle arrest, and enhanced programmed cell death along with augmentation in the level of ROS, dissipated mitochondrial membrane potential, activation of caspase cascade, and eventually inhibited the HH signaling cascade. In addition, CAR treatment increased the expression of pro-apoptotic proteins (Bax, Bad, Fas-L, TRAIL, FADDR, cytochrome c) and concomitantly reduced the expression of anti-apoptotic proteins (Bcl-2 and Bcl-xL) in C33A cells. CAR mediates the activation of caspase-9 and -3 (intrinsic pathway) and caspase-8 (extrinsic pathway) accompanied by the cleavage of PARP in cervical cancer cells. Thus, CAR induced apoptosis by both the intrinsic and extrinsic apoptotic pathways. CAR efficiently inhibited the growth of cervical cancer cells via arresting the cell cycle at G0/G1 phase and modulated the gene expression of related proteins (p21, p27, cyclin D1 and CDK4). Moreover, CAR inhibited the HH/GLI signaling pathway by down regulating the expression of SMO, PTCH and GLI1 proteins in cervical carcinoma cells. With evidence of the above results, our data revealed that CAR treatment suppressed the growth of HPV^-C33A cervical cancer cells and further elucidated the mechanistic insights into the functioning of CAR.

Keywords: ROS generation; anticancer; antioxidant activity; apoptosis; beta-glucan; cervical cancer.

Grants and funding

The authors are thankful to the Basic Science Research Program of the National Re-search Foundation of Korea (NRF) funded by the Ministry of Education (2021R1A6A1A03038785 and 2020R1I1A1A01073071).