Bafilomycin C1 induces G0/G1 cell-cycle arrest and mitochondrial-mediated apoptosis in human hepatocellular cancer SMMC7721 cells

Abstract

Bafilomycin C1, which was isolated from Streptomyces albolongus in our previous work, exhibited strong cytotoxicity against several cancer cell lines. This study aimed to evaluate its antitumor effect on human hepatocellular cancer SMMC7721 cells and the underlying mechanism in vitro and in vivo. MTT assay revealed that bafilomycin C1 retarded SMMC7721 cell growth and proliferation. Western blot and real-time qPCR analysis revealed that bafilomycin C1 caused partial G0/G1 phase cell-cycle arrest, downregulated the expression of cyclin D3, cyclin E1, CDK2, CDK4, and CDK6 and upregulated the expression of p21. Moreover, bafilomycin C1 caused mitochondrial membrane dysfunction through oxidative stress. Furthermore, bafilomycin C1 decreased the expression of Bcl-2; increased the expression of Bax, p53, and P-p53; and increased cleavage of caspase-9 and caspase-3, thereby inducing the intrinsic caspase-dependent apoptotic pathway. In vivo experiments in mice suggested that bafilomycin C1 suppressed tumor growth with few side effects. Cell-cycle arrest and induced apoptosis in tumor tissues in a mouse model treated with bafilomycin C1 were demonstrated by histological analyses, western blot and TUNEL. These findings indicate that bafilomycin C1 may be a promising candidate for hepatic cellular cancer therapy.

A semisynthetic borrelidin analogue BN-3b exerts potent antifungal activity against Candida albicans through ROS-mediated oxidative damage

“…The above results confirmed that BN-3b stimulated ROS accumulation in C. albicans. The excessive ROS production can lead to mitochondrial dysfunction, mainly characterized by the loss of MMP and decrease of ATP generation 28 . Our current study showed that the MMP levels of C. albicans cells were significantly reduced in a dose-dependent manner after treated with BN-3b.…”

“…BN-3b, AMB (Sigma-Aldrich) and FLC (Solarbio Science & Technology Co., Ltd., Beijing, China) were dissolved in dimethyl sulphoxide (DMSO) to prepare 150.0 mg/mL, 10.0 mg/mL and 10.0 mg/mL of stock solutions and stored at −20 °C. Working solutions of BN-3b, AMB, and FLC were prepared by diluting the stock solution with DMSO prior to use 28 . N-acetylcysteine (Beyotime Biotechnology Co., Shanghai, China) was dissolved in phosphate-buffered saline (PBS) to prepare 0.5 M of stock solution and stored at −20 °C.…”

“…C. albicans SC5314 (6 × 10 5 CFU/mL) incubated with vehicle or different concentrations of BN-3b (25.0, 50.0, and 75.0 µg/mL) for 8 h. Then cells (3 × 10 7 ) were incubated with a JC-1 staining solution at 37 °C for 20 min in the dark, washed twice with PBS, and resuspended in the buffer 28 . Green (excitation/emission wavelength: 514/529 nm) and red (excitation/emission wavelength: 585/590 nm) fluorescence were detected on a microplate reader 28 . The ratio of red/green fluorescence intensity represents MMP 22 .…”

Role of cyclin‐dependent kinases (CDKs) in hepatocellular carcinoma: Therapeutic potential of targeting the CDK signaling pathway

“…Additionally, by increasing p21 and p53 levels, mRNA and protein expression is notably reduced. Hepatocellular carcinoma cells are induced by bafilomycin C1 partial cell‐cycle arrest and activated apoptotic pathway both in vitro and in vivo , together with decreasing mRNA and protein levels of CDK2, increasing p21 and p53 …”

“…Hepatocellular carcinoma cells are induced by bafilomycin C1 partial cell- cycle arrest and activated apoptotic pathway both in vitro and in vivo, together with decreasing mRNA and protein levels of CDK2, increasing p21 and p53. 57 UCN-01 is also known as 7-hydroxystaurosporine and functions as a kinase inhibitor. UCN-01 inhibits checkpoint kinase 1 (CHK1), CHK2, protein kinase A, and protein kinase C, in addition to CDK2.…”