Abstract 【Objective】To investigate the potential effects of anlotinib on the growth of hepatocellular carcinoma (HCC) xenografts in nude mice,and its influences on the circadian (biological clock) gene expression in HCC tissues. 【Methods】We chose 36 athymic nude mice(half male and half female) for the study. The mice were divided randomly into two groups (the observation group and the control group), which were 18 mice per group. HepG2 cells were transplanted under the left axilla of the mice to establish the subcutaneous hepatocellular carcinoma xenograft model . After one week of HepG2 cell implatation, the observation group mice received anlotinib 50mg/(kg·d) for two weeks by intragastric administration, while the control group mice were given vehicle DMSO 100μL/20(g·d) in the same way gavage. Tumor volume was measured before and 24h after gavage. It was measured in every 2 days until experiment ended. In the end, the mice were sacrificed and tumors were isolated. Real-time PCR (rt-PCR) method was used to detect the mRNA level of circadian clock genes Per (Per1, Per2, Per3), CLOCK, Cry (Cry1, Cry2), Bmal1, and Dec2. At the same time we used western blot to evaluate the proteins of circadian genes expression. 【Results】 Compared to the control group, the xenograft tumors of the nude mice in the Anlotinib group grew slowly and tended to shrink in size, which were statistically significant (P<0.05). The results of rt-PCR indicated that the mRNA expressions of Per1 and Per2 in HCC cells were significantly increased under the action of anlotinib (P<0.05), while the downstream proteins increased by western blot. Meanwhile, the mRNA level of Dec2 were significantly inhibited (P<0.05) which accompanied with its downstream protein decreased. Though the mRNA level of Cry1 remained unchanged, however, the downstream protein expression of Cry1 was inhibited. 【Conclusion】Anlotinib can significantly regulate the expressions of various biological clock genes in HCC cells, which thereafter regulate their corresponding protein expression. Thus, anlotinib can significantly inhibit the tumor growth of HCC xenografts.
CHEN Zhuo-ying,WANG Yan,LIU Wen-sheng, et al. Effects of Anlotinib on Tumor Growth and Circadian Gene Expression of Hepatocellular Carcinoma Xenografts in Nude Mice[J]. JOURNAL OF CLINICAL RESEARCH, 2021, 38(1): 68-71.
[1] Torre LA, Bray F, Siegel RL, et al.Global cancer statistics, 2012[J]. CA Cancer J Clin,2015, 65(2): 87-108. [2] Lin B, Song X, Yang D, et al.Anlotinib inhibits angiogenesis via suppressing the activation of VEGFR2, PDGFRbeta and FGFR1[J].Gene,2018, 654: 77-86. [3] Khapre RV, Samsa WE, Kondratov RV.Circadian regulation of cell cycle: Molecular connections between aging and the circadian clock[J].Ann Med,2010, 42(6): 404-415. [4] Loudon, AS.Circadian biology: a 2.5 billion year old clock[J].Curr Biol,2012, 22(14): R570-R571. [5] Wu Y, Tang D, Liu N, et al.Reciprocal regulation between the circadian clock and hypoxia signaling at the genome level in mammals[J].Cell Metab,2017, 25(1): 73-85. [6] Gao Y, Liu P, Shi R.Anlotinib as a molecular targeted therapy for tumors[J].Oncol Lett,2020, 20(2): 1001-1014. [7] Takahashi JS.Transcriptional architecture of the mammalian circadian clock[J].Nat Rev Genet,2017, 18(3): 164-179. [8] Vollmers C, Gill S, Ditacchio L, et al.Time of feeding and the intrinsic circadian clock drive rhythms in hepatic gene expression[J].Proc Natl Acad Sci USA,2009, 106(50): 21453-21458. [9] Mazzoccoli G, Miele L, Marrone G, et al.A role for the biological clock in liver cancer[J].Cancers (Basel),2019, 11(11):1778. [10] Liu B, Xu k, Jiang Y, et al.Aberrant expression of Per1, Per2 and Per3 and their prognostic relevance in non-small cell lung cancer[J].Int J Clin Exp Pathol,2014, 7(11): 7863-7871. [11] 张建省,袁鹏,颜昭勇,等. 节律分子Timeless调控糖代谢重编程促进肝癌细胞生长[J].中华肿瘤杂志, 2018, 40(7): 499-505.
2021, Vol. 38 
