Abstract��OBJECTIVE To investigate the apoptosis effect of human hepatocellular carcinoma cell line SMMC-7721 induced by dihydroartemisinin in vitro and the possible mechanism. METHODS After treatment with 25, 50, 100, 200, and 400 ��mol��L-1 dihydroartemisinin for 24 h. The proliferation inhibitory effect of dihydroartemisinin on SMMC-7721 cell was detected by MTT assay. Cell cycle and apoptosis were detected by flow cytometry. The change of apoptotic morphology was detected by confocal laser scanning microscopy. Rho 123 staining method was used to detect the changes of mitochondrial membrane potential. Western blot was used to detect expression of Bcl-2, Bax, Cleaved Caspase-3, Cleaved Caspase-9 and Cyto C. RESULTS MTT results showed that 25-400 ��mol��L-1 dihydroartemisinin can inhibit the proliferation of SMMC-7721 cells obviously. The cell cycle detection results of flow cytometry showed that dihydroartemisinin could block SMMC-7721 cell cycle in G2/M phase. The results of Hochest 333258 staining showed that the nuclei were heterogeneous, condensed and fragmented in the DHA treatment group. The cell apoptosis detection results of flow cytometry showed that the apoptosis rate of dihydroartemisinin treated groups were increased obviously (P<0.01). The results of Rho 123 staining showed that the mitochondrial membrane potential was decreased significantly (P<0.01). Western blot results showed that the expression of Bcl-2 was down-regulated, expression of Bax was up-regulated, the ration of Bax/Bcl-2 was increased and the expression of Cleaved Caspase-3, Cleaved Caspase-9 and Cyto C were up-regulated. CONCLUSION Dihydroartemisinin can induce apoptosis of SMMC-7721 cells, on the mechanism of apoptosis may be related to mitochondrial pathway.
JIAO Z, LING G, XIA Z, et al. Dihydroartemisinin induces endothelial cell anoikis through the activation of the JNK signaling pathway [J]. Oncol Lett, 2016, 12(3): 1896-1900.
[2]
LIN R, ZHANG Z, CHEN L, et al. Dihydroartemisinin (DHA) induces ferroptosis and causes cell cycle arrest in head and neck carcinoma cells [J]. Cancer Lett, 2016, 381(1): 165-175.
[3]
RIJKEN M J, MCGREADY R, BOEL M E, et al. Dihydroartemisinin-piperaquine rescue treatment of multidrug-resistant Plasmodium falciparum malaria in pregnancy: a preliminary report [J]. Am J Trop Med Hyg, 2016, 78(4): 543-545.
[4]
CHEN W Q, QI H W, WU C G, et al. Inhibition on the human lung adenocarcinoma cell line A549 and A549/CDDP in vitro by dihydroartemisini in combination with cisplatin [J]. Chin J Clin Rehabil(�й��ٴ�����), 2005, 43(9): 120-122.
[5]
LU J J, HUANG M X, CHEN X P, et al. Progress in anti-neoplastic effects and mechanisms of dihydroartemisinin [J]. Chin J Pharmacol Toxicol(�й�ҩ��ѧ�붾��ѧ��־), 2012, 26 (3):397-401.
[6]
MEHDIZADEH A, SOMI M H, DARABI M, et al. Extracellular signal-regulated kinase 1 and 2 in cancer therapy: a focus on hepatocellular carcinoma [J]. Mol Biol Reports, 2016, 43(2): 107-116.
[7]
CHEN X, HAN Y, ZHAN S, et al. Tegillarca granosa extract haishengsu induces apoptosis in human hepatocellular carcinoma cell line BEL-7402 via fas-signaling pathways [J]. Cell Biochem Biophys, 2015, 71(2): 837-844.
[8]
HU F, HAN J, ZHAI B, et al. Blocking autophagy enhances the apoptosis effect of bufalin on human hepatocellular carcinoma cells through endoplasmic reticulum stress and JNK activation [J]. Apoptosis, 2014, 19(1): 210-223.
[9]
XU C C, DENG T, FAN M L, et al. Synthesis and in vitro antitumor evaluation of dihydroartemisinin-cinnamic acid ester derivatives [J]. Euro J Med Chem, 2016, 107:192-203.
[10]
BOWARD B, WU T, DALTON S. Concise review: control of cell fate through cell cycle and pluripotency networks [J]. Stem Cells, 2016, 34(6): 1427.
[11]
KASTAN, AMP M B, BARTEK J. Cell-cycle checkpoints and cancer [J]. Nature, 2004, 432(7015): 316-323.
[12]
MATT S, HOFMANN T G. The DNA damage-induced cell death response: a roadmap to kill cancer cells [J]. Cell Mol Life Sci Cmls, 2016, 73(15): 2829-2850.
[13]
MA R, YANG L, NIU F, et al. HIV tat-mediated induction of human brain microvascular endothelial cell apoptosis involves endoplasmic reticulum stress and mitochondrial dysfunction [J]. Mol Neurobiol, 2016, 53(1): 132-142.
[14]
SIDDIQUI W A, AHAD A, AHSAN H. The mystery of BCL2 family: Bcl-2 proteins and apoptosis: an update [J]. Arch Toxicol, 2015, 89(3): 289-317.
[15]
WANG X, YAN Y, YANG L, et al. Effect of quercetin on the expression of Bcl-2/Bax apoptotic proteins in endometrial cells of lipopolysaccharide-induced-abortion mice [J]. J Tradit Chin Med(��ҽ��־), 2016, 36(6): 737-742.
[16]
ZHU W, ZHANG W, WANG H, et al. Apoptosis induced by microwave radiation in pancreatic cancer JF305 cells [J]. Can J Physiol Pharmacol, 2014, 92(4): 324-329.
[17]
FANG H H, LI H J, KE X L, et al. Recent research advances in anti-tumor effects of berberine and its derivatives [J]. Chin Pharm J (�й�ҩѧ��־), 2017, 52(6): 429-433.
[18]
ZOU X, QU Z Y, JI Y B. Juglone induces apoptosis of human gastric cancer SGC-7901 cells through reactive oxygen species mediated JNK and p38 pathway [J]. Chin Pharm J (�й�ҩѧ��־), 2016, 51(7): 544-549.