PMID- 37802809 OWN - NLM STAT- MEDLINE DCOM- 20231101 LR - 20231101 IS - 1001-5302 (Print) IS - 1001-5302 (Linking) VI - 48 IP - 17 DP - 2023 Sep TI - [Inhibitory effect and molecular mechanism of sinomenine on human hepatocellular carcinoma HepG2 and SK-HEP-1 cells]. PG - 4702-4710 LID - 10.19540/j.cnki.cjcmm.20230424.401 [doi] AB - This study aimed to investigate the effect and molecular mechanism of sinomenine on proliferation, apoptosis, metastasis, and combination with inhibitors in human hepatocellular carcinoma HepG2 cells and SK-HEP-1 cells. The effect of sinomenine on the growth ability of HepG2 and SK-HEP-1 cells were investigated by CCK-8 assay, colony formation assay, and BeyoClick~(TM) EdU-488 staining. The effect of sinomenine on DNA damage was detected by immunofluorescence assay, and the effect of sinomenine on apoptosis of human hepatocellular carcinoma cells was clarified by Hoechst 33258 staining and CellEvent~(TM) Cystein-3/7Green ReadyProbes~(TM) reagent assay. Cell invasion assay and 3D tumor cell spheroid invasion assay were performed to investigate the effect of sinomenine on the invasion ability of human hepatocellular carcinoma cells in vitro. The effect of sinomenine on the regulation of protein expression related to the protein kinase B(Akt)/mammalian target of rapamycin(mTOR)/signal transducer and activator of transcription 3(STAT3) signaling pathway in HepG2 and SK-HEP-1 cells was examined by Western blot. Molecular docking was used to evaluate the strength of affinity of sinomenine to the target cysteinyl aspartate specific proteinase-3(caspase-3) and STAT3, and combined with CCK-8 assay to detect the changes in cell viability after combination with STAT3 inhibitor JSI-124 in combination with CCK-8 assay. The results showed that sinomenine could significantly reduce the cell viability of human hepatocellular carcinoma cells in a concentration-and time-dependent manner, significantly inhibit the clonogenic ability of human hepatocellular carcinoma cells, and weaken the invasive ability of human hepatocellular carcinoma cells in vitro. In addition, sinomenine could up-regulate the cleaved level of poly ADP-ribose polymerase(PARP), a marker of apoptosis, and down-regulate the protein levels of p-Akt, p-mTOR, and p-STAT3 in human hepatocellular carcinoma cells. Molecular docking results showed that sinomenine had good affinity with the targets caspase-3 and STAT3, and the sensitivity of sinomenine to hepatocellular carcinoma cells was diminished after STAT3 was inhibited. Therefore, sinomenine can inhibit the proliferation and invasion of human hepatocellular carcinoma cells and induce apoptosis, and the mechanism may be attributed to the activation of caspase-3 signaling and inhibition of the Akt/mTOR/STAT3 pathway. This study can provide a new reference for the in-depth research and clinical application of sinomenine and is of great significance to further promote the scientific development and utilization of sinomenine. FAU - Tian, Ying-Ying AU - Tian YY AD - School of Chinese Materia Medica,Beijing University of Chinese Medicine Beijing 100029,China. FAU - Ma, Bei-Bei AU - Ma BB AD - Institute of Scientific Research, Beijing Tong Ren Tang Chinese Medicine Co., Ltd. Beijing 100079,China. FAU - Zhao, Xin-Yue AU - Zhao XY AD - School of Chinese Materia Medica,Beijing University of Chinese Medicine Beijing 100029,China. FAU - Liu, Chuang AU - Liu C AD - School of Chinese Materia Medica,Beijing University of Chinese Medicine Beijing 100029,China. FAU - Li, Yi-Lin AU - Li YL AD - School of Chinese Materia Medica,Beijing University of Chinese Medicine Beijing 100029,China. FAU - Yu, Shang-Yue AU - Yu SY AD - School of Chinese Materia Medica,Beijing University of Chinese Medicine Beijing 100029,China. FAU - Tian, Shi-Qiu AU - Tian SQ AD - School of Chinese Materia Medica,Beijing University of Chinese Medicine Beijing 100029,China. FAU - Pei, Hai-Luan AU - Pei HL AD - School of Chinese Materia Medica,Beijing University of Chinese Medicine Beijing 100029,China. FAU - Lyu, Ying-Nan AU - Lyu YN AD - School of Chinese Materia Medica,Beijing University of Chinese Medicine Beijing 100029,China. FAU - Zuo, Ze-Ping AU - Zuo ZP AD - Institute of Scientific Research, Beijing Tong Ren Tang Chinese Medicine Co., Ltd. Beijing 100079,China. FAU - Wang, Zhi-Bin AU - Wang ZB AD - School of Chinese Materia Medica,Beijing University of Chinese Medicine Beijing 100029,China Institute of Scientific Research, Beijing Tong Ren Tang Chinese Medicine Co., Ltd. Beijing 100079,China. LA - chi PT - English Abstract PT - Journal Article PL - China TA - Zhongguo Zhong Yao Za Zhi JT - Zhongguo Zhong yao za zhi = Zhongguo zhongyao zazhi = China journal of Chinese materia medica JID - 8913656 RN - EC 2.7.11.1 (Proto-Oncogene Proteins c-akt) RN - EC 3.4.22.- (Caspase 3) RN - 63LT81K70N (sinomenine) RN - M03GIQ7Z6P (Sincalide) RN - EC 2.7.11.1 (TOR Serine-Threonine Kinases) SB - IM MH - Humans MH - *Carcinoma, Hepatocellular/drug therapy/genetics MH - Proto-Oncogene Proteins c-akt/metabolism MH - Caspase 3/metabolism MH - *Liver Neoplasms/drug therapy/genetics MH - Molecular Docking Simulation MH - Sincalide/pharmacology MH - Cell Line, Tumor MH - Cell Proliferation MH - Hep G2 Cells MH - TOR Serine-Threonine Kinases/metabolism MH - Apoptosis OTO - NOTNLM OT - Akt/mTOR/STAT3 signaling pathway OT - apoptosis OT - caspase-3 OT - hepatocellular carcinoma OT - invasion OT - molecular docking OT - sinomenine EDAT- 2023/10/07 00:41 MHDA- 2023/11/01 12:44 CRDT- 2023/10/06 22:03 PHST- 2023/11/01 12:44 [medline] PHST- 2023/10/07 00:41 [pubmed] PHST- 2023/10/06 22:03 [entrez] AID - 10.19540/j.cnki.cjcmm.20230424.401 [doi] PST - ppublish SO - Zhongguo Zhong Yao Za Zhi. 2023 Sep;48(17):4702-4710. doi: 10.19540/j.cnki.cjcmm.20230424.401.