Circadian regulator BMAL1::CLOCK promotes cell proliferation in hepatocellular carcinoma by controlling apoptosis and cell cycle

Hepatocellular carcinoma (HCC) remains a global health challenge whose incidence is growing worldwide. Previous evidence strongly supported the notion that the circadian clock controls physiological homeostasis of the liver and plays a key role in hepatocarcinogenesis. Despite the progress, cellular and molecular mechanisms underpinning this HCC-clock crosstalk remain unknown. Addressing this knowledge gap, we show here that although the human HCC cells Hep3B, HepG2, and Huh7 displayed variations in circadian rhythm profiles, all cells relied on the master circadian clock transcription factors, BMAL1 and CLOCK, for sustained cell growth. Down-regulating Bmal1 or Clock in the HCC cells induced apoptosis and arrested cell cycle at the G2/M phase. Mechanistically, we found that inhibiting Bmal1/Clock induced dysregulation of the cell cycle regulators Wee1 and p21 which cooperatively contribute to tumor cell death. Bmal1/Clock knockdown caused downregulation of Wee1 that led to apoptosis activation and upregulation of p21 which arrested the cell cycle at the G(2)/M phase. Collectively, our results suggest that the circadian clock regulators BMAL1 and CLOCK promote HCC cell proliferation by controlling Wee1 and p21 levels, thereby preventing apoptosis and cell cycle arrest. Our findings shed light on cellular impact of the clock proteins for maintaining HCC oncogenesis and provide proof-of-principle for developing cancer therapy based on modulation of the circadian clock.

Automated summary

Hepatocellular carcinoma (HCC) is a global health challenge with increasing incidence worldwide. The circadian clock has been shown to play a key role in hepatocarcinogenesis, but the cellular and molecular mechanisms underlying the HCC-clock crosstalk are unknown. In this study, we found that HCC cells rely on the circadian clock transcription factors BMAL1 and CLOCK for cell growth, and down-regulation of these factors induces apoptosis and cell cycle arrest. Mechanistically, inhibiting BMAL1/CLOCK dysregulates cell cycle regulators Wee1 and p21, leading to tumor cell death. Our findings provide insights into the cellular impact of clock proteins in HCC oncogenesis and suggest a potential therapeutic approach based on modulation of the circadian clock.