TY - JOUR
T1 - Aloperine induces apoptosis and G2/M cell cycle arrest in hepatocellular carcinoma cells through the PI3K/Akt signaling pathway
AU - Liu, Jun Shan
AU - Huo, Chu Ying
AU - Cao, Hui Hui
AU - Fan, Chun Lin
AU - Hu, Jian Yang
AU - Deng, Li Juan
AU - Lu, Zi Bin
AU - Yang, Hua Yi
AU - Yu, Lin Zhong
AU - Mo, Zhi Xian
AU - YU, Zhiling
N1 - Funding Information:
This work was supported by the National Natural Science Foundation of China ( 81402801 , 81602997 , 81803790 ), the Guangdong Province Universities and Colleges Pearl River Scholar Funded Scheme ( GDHVPS2018 ), the Guangdong National Natural Science Funds for Distinguished Young Scholar ( 2017A030306006 ), Medical Scientific Research Foundation of Guangdong Province ( 2017373 ), the Program of Pearl River Young Talents of Science and Technology in Guangzhou , China ( 201710010026 ) and the Hong Kong Scholars program ( XJ2016059 ).
PY - 2019/8
Y1 - 2019/8
N2 - Background: Hepatocellular carcinoma (HCC)ranks third among the most common causes of cancer-related deaths worldwide. The chemotherapy for HCC is still insufficient, so far. In searching for effective anti-HCC agents from traditional Chinese medicine, we discovered that aloperine (ALO), a quinolizidine alkaloid from Sophora alopecuroides L., exerts anti-HCC activities. However, the effects of ALO on HCC have been rarely studied, and its underlying mechanisms remain unknown. Purpose: This study aims to evaluate the anti-HCC activities of ALO and explore its underlying mechanisms. Methods: MTT assay and colony formation assay were used to investigate the anti-proliferative effects of ALO on human HCC Hep3B and Huh7 cells. Hoechst 33258 staining was used to observe the morphological changes of cells after ALO treatment. Flow cytometry was used to analyze apoptosis induction, the collapse of the mitochondrial membrane potential and cell cycle distribution. Western blotting was used to examine the expression levels of proteins associated with apoptosis and cell cycle arrest, and key proteins in the PI3K/Akt signaling pathway. Small interfering RNA (siRNA)transfection was used to investigate the role of Akt in ALO-induced apoptosis and cell cycle arrest. Zebrafish tumor model was used to evaluate the anti-HCC effects of ALO in vivo. Results: ALO inhibited the proliferation of Hep3B and Huh7 cells. ALO induced apoptosis in HCC cells, which was accompanied by the loss of mitochondrial potential, the release of cytochrome c into cytosol, as well as the increased cleavages of caspase-9, caspase-3 and PARP. Moreover, ALO induced G2/M cell cycle arrest by downregulating the expression levels of cdc25C, cdc2 and cyclin B1. In addition, ALO inhibited activation of the PI3K/Akt signaling pathway by decreasing the expression levels of p110α, p85, Akt and p-Akt (Ser473). Further study showed that inhibition of Akt by siRNA augmented ALO-mediated apoptosis and G2/M cell cycle arrest in HCC cells. Critically, ALO inhibited the growth of Huh7 cells in vivo. Conclusion: We first demonstrated that ALO induced apoptosis and G2/M cell cycle arrest in HCC cells through inhibition of the PI3K/Akt signaling pathway. This study provides a rationale for ALO as a potential chemotherapeutic agent for HCC.
AB - Background: Hepatocellular carcinoma (HCC)ranks third among the most common causes of cancer-related deaths worldwide. The chemotherapy for HCC is still insufficient, so far. In searching for effective anti-HCC agents from traditional Chinese medicine, we discovered that aloperine (ALO), a quinolizidine alkaloid from Sophora alopecuroides L., exerts anti-HCC activities. However, the effects of ALO on HCC have been rarely studied, and its underlying mechanisms remain unknown. Purpose: This study aims to evaluate the anti-HCC activities of ALO and explore its underlying mechanisms. Methods: MTT assay and colony formation assay were used to investigate the anti-proliferative effects of ALO on human HCC Hep3B and Huh7 cells. Hoechst 33258 staining was used to observe the morphological changes of cells after ALO treatment. Flow cytometry was used to analyze apoptosis induction, the collapse of the mitochondrial membrane potential and cell cycle distribution. Western blotting was used to examine the expression levels of proteins associated with apoptosis and cell cycle arrest, and key proteins in the PI3K/Akt signaling pathway. Small interfering RNA (siRNA)transfection was used to investigate the role of Akt in ALO-induced apoptosis and cell cycle arrest. Zebrafish tumor model was used to evaluate the anti-HCC effects of ALO in vivo. Results: ALO inhibited the proliferation of Hep3B and Huh7 cells. ALO induced apoptosis in HCC cells, which was accompanied by the loss of mitochondrial potential, the release of cytochrome c into cytosol, as well as the increased cleavages of caspase-9, caspase-3 and PARP. Moreover, ALO induced G2/M cell cycle arrest by downregulating the expression levels of cdc25C, cdc2 and cyclin B1. In addition, ALO inhibited activation of the PI3K/Akt signaling pathway by decreasing the expression levels of p110α, p85, Akt and p-Akt (Ser473). Further study showed that inhibition of Akt by siRNA augmented ALO-mediated apoptosis and G2/M cell cycle arrest in HCC cells. Critically, ALO inhibited the growth of Huh7 cells in vivo. Conclusion: We first demonstrated that ALO induced apoptosis and G2/M cell cycle arrest in HCC cells through inhibition of the PI3K/Akt signaling pathway. This study provides a rationale for ALO as a potential chemotherapeutic agent for HCC.
KW - Aloperine
KW - Apoptosis
KW - G2/M cell cycle arrest
KW - Hepatocellular carcinoma
KW - PI3K/Akt signaling pathway
UR - http://www.scopus.com/inward/record.url?scp=85064731237&partnerID=8YFLogxK
U2 - 10.1016/j.phymed.2019.152843
DO - 10.1016/j.phymed.2019.152843
M3 - Journal article
C2 - 31039533
AN - SCOPUS:85064731237
SN - 0944-7113
VL - 61
JO - Phytomedicine
JF - Phytomedicine
M1 - 152843
ER -
