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Though Ginsenoside F2 (GF2), a protopanaxadiol saponin from Panax ginseng, is known to have an anticancer effect, its underlying mechanism still remains unclear. In our model, the anti-glycolytic mechanism of GF2 was investigated in human cervical cancer cells in association with miR193a-5p and the β-catenin/c-Myc/Hexokinase 2 (HK2) signaling axis. Here, GF2 exerted significant cytotoxicity and antiproliferation activity, increased sub-G1, and attenuated the expression of pro-Poly (ADPribose) polymerase (pro-PARP) and pro-cysteine aspartyl-specific protease (procaspase3) in HeLa and SiHa cells. Consistently, GF2 attenuated the expression of Wnt, β-catenin, and c-Myc and their downstream target genes such as HK2, pyruvate kinase isozymes M2 (PKM2), and lactate dehydrogenase A (LDHA), along with a decreased production of glucose and lactate in HeLa and SiHa cells. Moreover, GF2 suppressed β-catenin and c-Myc stability in the presence and absence of cycloheximide in HeLa cells, respectively. Additionally, the depletion of β-catenin reduced the expression of c-Myc and HK2 in HeLa cells, while pyruvate treatment reversed the ability of GF2 to inhibit β-catenin, c-Myc, and PKM2 in GF2-treated HeLa cells. Notably, GF2 upregulated the expression of microRNA139a-5p (miR139a-5p) in HeLa cells. Consistently, the miR139a-5p mimic enhanced the suppression of β-catenin, c-Myc, and HK2, while the miR193a-5p inhibitor reversed the ability of GF2 to attenuate the expression of β-catenin, c-Myc, and HK2 in HeLa cells. Overall, these findings suggest that GF2 induces apoptosis via the activation of miR193a-5p and the inhibition of β-catenin/c-Myc/HK signaling in cervical cancer cells. [ABSTRACT FROM AUTHOR]

Though cornin is known to induce angiogenic, cardioprotective, and apoptotic effects, the apoptotic mechanism of this iridoid monoglucoside is not fully understood in prostate cancer cells to date. To elucidate the antitumor mechanism of cornin, cytotoxicity assay, cell cycle analysis, Western blotting, RT‐qPCR, RNA interference, immunofluorescence, immunoprecipitation, reactive oxygen species (ROS) measurement, and inhibitor assay were applied in this work. Cornin exerted cytotoxicity, increased sub‐G1 population, and cleaved PARP and caspase3 in LNCaP cells more than in DU145 cells. Consistently, cornin suppressed phosphorylation of signal transducer and activator of transcription 3 (STAT3) and disrupted the colocalization of STAT3 and androgen receptor (AR) in LNCaP and DU145 cells, along with suppression of AR, prostate‐specific antigen (PSA), and 5α‐reductase in LNCaP cells. Furthermore, cornin increased ROS production and the level of miR‐193a‐5p, while ROS inhibitor N‐acetylcysteine disturbed the ability of cornin to attenuate the expression of AR, p‐STAT3, PSA, pro‐PARP, and pro‐caspase3 in LNCaP cells. Notably, miR‐193a‐5p mimics the enhanced apoptotic effect of cornin, while miR‐193a‐5p inhibitor reverses the ability of cornin to abrogate AR, PSA, and STAT3 in LNCaP cells. Our findings suggest that ROS production and the disturbed crosstalk between STAT3 and AR by microRNA‐193a‐5p are critically involved in the apoptotic effect of cornin in prostate cancer cells. [ABSTRACT FROM AUTHOR]

Though Morusin is known to induce apoptotic, antiprolifertaive, and autophagic effects through several signaling pathways, the underlying molecular mechanisms of Morusin still remain unclear until now. To elucidate antitumor mechanism of Morusin, cytotoxicity assay, cell cycle analysis, Western blotting, TUNEL assay, RNA interference, immunofluorescense, immunoprecipitation, reactive oxygen species (ROS) measurement, and inhibitor study were applied in this study. Morusin enhanced cytotoxicity, increased the number of TUNEL positive cells, sub‐G1 population and induced the cleavages of PARP and caspase3, attenuated the expression of HK2, PKM2, LDH, c‐Myc, and Forkhead Box M1 (FOXM1) along with the reduction of glucose, lactate, and ATP in DU145 and PC3 cells. Furthermore, Morusin disrupted the binding of c‐Myc and FOXM1 in PC‐3 cells, which was supported by String and cBioportal database. Notably, Morusin induced c‐Myc degradation mediated by FBW7 and suppressed c‐Myc stability in PC3 cells exposed to MG132 and cycloheximide. Also, Morusin generated ROS, while NAC disrupted the capacity of Morusin to reduce the expression of FOXM1, c‐Myc, pro‐PARP, and pro‐caspase3 in PC‐3 cells. Taken together, these findings provide scientific evidence that ROS mediated inhibition of FOXM1/c‐Myc signaling axis plays a critical role in Morusin induced apoptotic and anti‐Warburg effect in prostate cancer cells. Our findings support scientific evidence that ROS mediated inhibition of FOXM1/c‐Myc signaling axis is critically involved in apoptotic and anti‐Warburg effect of Morusin in prostate cancer cells. [ABSTRACT FROM AUTHOR]

To elucidate the underlying antitumor mechanism of lambertianic acid (LA) derived from Pinus koraiensis, the role of cancer metabolism related molecules was investigated in the apoptotic effect of LA in DU145 and PC3 prostate cancer cells. MTT assay for cytotoxicity, RNA interference, cell cycle analysis for sub G1 population, nuclear and cytoplasmic extraction, lactate, Glucose and ATP assay by ELISA, Measurement of reactive oxygen species (ROS) generation, Western blotting, and immunoprecipitation assay were conducted in DU145 and PC3 prostate cancer cells. Herein LA exerted cytotoxicity, increased sub G1 population and attenuated the expression of pro‐Caspase3 and pro‐poly (ADP‐ribose) polymerase (pro‐PARP) in DU145 and PC3 cells. Also, LA reduced the expression of lactate dehydrogenase A (LDHA), glycolytic enzymes such as hexokinase 2 and pyruvate kinase M2 (PKM2) with reduced production of lactate in DU145 and PC3 cells. Notably, LA decreased phosphorylation of PKM2 on Tyr105 and inhibited the expression of p‐STAT3, cyclin D1, C‐Myc, β‐catenin, and p‐GSK3β with the decrease of nuclear translocation of p‐PKM2. Furthermore, LA disturbed the binding of p‐PKM2 and β‐catenin in DU145 cells, which was supported by Spearman coefficient (0.0463) of cBioportal database. Furthermore, LA generated ROS in DU145 and PC3 cells, while ROS scavenger NAC (N‐acetyl L‐cysteine) blocked the ability of LA to reduce p‐PKM2, PKM2, β‐catenin, LDHA, and pro‐caspase3 in DU145 cells. Taken together, these findings provide evidence that LA induces apoptosis via ROS generation and inhibition of PKM2/β‐catenin signaling in prostate cancer cells. [ABSTRACT FROM AUTHOR]

Though Honokiol was known to have anti‐inflammatory, antioxidant, anticancer, antithrombotic, anti‐viral, metabolic, antithrombotic, and neurotrophic activities, the underlying mechanisms of Honokiol on epithelial–mesenchymal transition (EMT) mediated liver fibrosis still remain elusive so far. Anti‐EMT and antifibrotic effects of Honokiol were explored in murine AML‐12 hepatocyte cells by 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyl‐2H‐tetrazolium bromide (MTT) assay, wound healing assay, Western blotting and also in CCl4‐induced liver injury mouse model by immunohistochemistry. Honokiol significantly suppressed transforming growth factor β1 (TGF‐β1)‐induced EMT and migration of AML‐12 cells along with decreased EMT phenotypes such as loss of cell adhesion and formation of fibroblast like mesenchymal cells in TGF‐β1‐treated AML‐12 cells. Consistently, Honokiol suppressed the expression of Snail and transmembrane protease serine 4 (TMPRSS4), but not p‐Smad3, and activated E‐cadherin in TGF‐β1‐treated AML‐12 cells. Additionally, Honokiol reduced the expression of β‐catenin, p‐AKT, p‐ERK, p‐p38 and increased phosphorylation of glycogen synthase kinase 3 beta (GSK3β) and JNK in TGF‐β1‐treated AML‐12 cells via TGF‐β1/nonSmad pathway. Conversely, GSK3β inhibitor SB216763 reversed the ability of Honokiol to reduce Snail, β‐catenin and migration and activate E‐cadherin in TGF‐β1‐treated AML‐12 cells. Also, Honokiol suppressed hepatic steatosis and necrosis by reducing the expression of TGF‐β1 and α‐SMA in liver tissues of CCl4 treated mice. These findings provide scientific evidence that Honokiol suppresses EMT and hepatic fibrosis via activation of E‐cadherin/GSK3β/JNK and inhibition of AKT/ERK/p38/β‐catenin/TMPRSS4 signaling axis. [ABSTRACT FROM AUTHOR]

The aim of the present study is to explore the underlying hepatoprotective mechanism of PKC#963, consisting of Pinus koraiensis, Saururus chinensis, and Lycium barbarum in association with acute and chronic liver injury induced by alcohol or carbon tetrachloride (CCl4). Here, PKC#963 significantly suppressed aspartate aminotransferase (AST), alanine aminotransferase (ALT), phosphorylated signal transducer and activator of transcription 3 (p‐STAT3), inducible nitric oxide synthase (iNOS), and cyclooxygenase (COX‐2) in CCl4‐treated HepG2 cells. Also, PKC#963 significantly suppressed reactive oxygen species (ROS) production in HepG2 cells. Consistently, PKC#963 suppressed the expression of AST, ALT, p‐STAT3, iNOS, COX‐2, interleukin 6 (IL‐6), tumor necrosis factor alpha (TNF‐α) and α‐smooth muscle actin (α‐SMA) and increased procaspase 3 in the liver tissues of CCl4 treated rats. In addition, PKC#963 enhanced alcohol dehydrogenase (ADH), aldehyde dehydrogenase (ALDH) for alcohol metabolism, superoxide dismutase (SOD), and catalase as antioxidant enzymes and also suppressed AST and ALT in alcohol‐treated rats. Furthermore, PKC#963 reduced hepatic steatosis and necrosis in CCl4‐treated rats by H&E (Hematoxylin and Eosin) staining. Taken together, these findings highlight evidence that PKC#963 has hepatoprotective potential via inhibition of iNOS, COX‐2, and p‐STAT3 and enhancement of SOD and catalase. [ABSTRACT FROM AUTHOR]

To target benign prostatic hyperplasia (BPH) as a common urinary disease in old men, in the current study, the antiproliferative and apoptotic mechanism of SH‐PRO, a mixture of Angelica gigas and Astragalus membranaceus (2:1), was evaluated in BPH‐1 cells and rats with testosterone‐induced BPH. Herein, SH‐PRO significantly reduced the viability of BPH‐1 cells and dihydrotestosterone (DHT)‐treated RWPE‐1 cells. Also, SH‐PRO increased the sub‐G1 population in BPH‐1 cells and consistently attenuated the expression of pro‐PARP, pro‐caspase 3, Bcl2, FOXO3a, androgen receptor (AR), and prostate‐specific antigen (PSA) in BPH‐1 cells and DHT‐treated RWPE‐1 cells. Of note, SH‐PRO generated reactive oxygen species (ROS) in BPH‐1 cells, while ROS inhibitor N‐acetyl‐l‐cysteine (NAC) disturbed the ability of SH‐PRO to reduce the expression of pro‐PARP, FOXO3a, catalase, SOD, and increase sub‐G1 population in BPH‐1 cells. Furthermore, oral treatment of SH‐PRO significantly abrogated the weight of the prostate in testosterone‐treated rats compared to BPH control with the reduced expression of AR, PSA, and DHT and lower plasma levels of DTH, bFGF, and EGF with no toxicity. Overall, these findings highlight the antiproliferative and apoptotic potential of SH‐PRO via ROS‐mediated activation of PARP and caspase 3 and inhibition of FOXO3a/AR/PSA signaling as a potent anti‐BPH candidate. [ABSTRACT FROM AUTHOR]

Herein, the apoptotic mechanism of 1,2,3,4,6-penta-O-galloyl-β-D-glucopyranose (PGG) was examined in cisplatin-resistant lung cancer cells. PGG significantly reduced viability; increased sub-G1 accumulation and the number of terminal deoxynucleotidyl transferase (TdT) dUTP Nick-End Labeling (TUNEL)-positive cells; induced the cleavage of poly (ADP-ribose) polymerase (PARP), caspases (8,9,3,7), B-cell lymphoma protein 2 (Bcl-2)-associated X (Bax) and phosphatase and tensin homolog deleted on chromosome 10 (PTEN); and attenuated the expression of p-AKT, X-linked inhibitor of apoptosis protein (XIAP), Bcl-2, Bcl-xL and survivin in A549/cisplatin-resistant (CR) and H460/CR cells. Notably, PGG activated p53, p-checkpoint kinase 2 (CHK2) and p-H2A histone family member X (p-H2AX), with increased levels of DNA damage (DSBs) evaluated by highly expressed pH2AX and DNA fragmentation registered on comet assay, while p53 knockdown reduced the ability of PGG to reduce viability and cleave caspase 3 and PARP in A549/CR and H460/CR cells. Additionally, PGG treatment suppressed the growth of H460/CR cells in Balb/c athymic nude mice with increased caspase 3 expression compared with the cisplatin group. Overall, PGG induces apoptosis in cisplatin-resistant lung cancer cells via the upregulation of DNA damage proteins such as γ-H2AX, pCHK2 and p53. [ABSTRACT FROM AUTHOR]

Herein, apoptotic mechanism of Moracin D was explored in prostate cancer cells in association with peroxisome proliferator‐activated receptor gamma (PPAR‐γ)‐related signaling involved in lipid metabolism. Moracin D augmented cytotoxicity and sub G1 population in PC3 and DU145 prostate cancer cells, while DU145 cells were more susceptible to Moracin D than PC3 cells. Moracin D attenuated the expression of caspase‐3, poly (ADP‐ribose) polymerase (PARP), B‐cell lymphoma 2 (Bcl‐2), and B‐cell lymphoma‐extra‐large (Bcl‐xL) in DU145 cells. Consistently, Moracin D significantly augmented the number of terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL)‐positive cells in DU145 cells. Interestingly, Moracin D activated PPAR‐γ and phospho‐protein kinase C delta (p‐PKC‐δ) and inhibited phospho‐protein kinase C alpha (p‐PKC‐α) in DU145 cells. Furthermore, STRING bioinformatic analysis reveals that PPAR‐γ interacts with nuclear factor‐κB (NF‐κB) that binds to PKC‐α/PKC‐δ or protein kinase B (AKT) or extracellular signal–regulated kinase (ERK). Indeed, Moracin D decreased phosphorylation of NF‐κB, ERK, and AKT in DU145 cells. Conversely, PPAR‐γ inhibitor GW9662 reduced the apoptotic ability of Moracin D to activate caspase 3 and PARP in DU145 cells. Taken together, these findings provide a novel insight that activation of PPAR‐γ/p‐PKC‐δ and inhibition of p‐PKC‐α are critically involved in Moracin D–induced apoptosis in DU145 prostate cancer cells. [ABSTRACT FROM AUTHOR]

Though Sanggenon G (SanG) from root bark of Morus alba was known to exhibit anti‐oxidant and anti‐depressant effects, its underlying mechanisms still remain unclear. Herein SanG reduced the viability of A549 and H1299 non‐small lung cancer cells (NSCLCs). Also, SanG increased sub‐G1 population via inhibition of cyclin D1, cyclin E, CDK2, CDK4 and Bcl‐2, cleavages of poly (ADP‐ribose) polymerase (PARP) and caspase‐3 in A549 and H1299 cells. Of note, SanG effectively inhibited c‐Myc expression by activating ribosomal protein L5 (RPL5) and reducing c‐Myc stability even in the presence of cycloheximide and 20% serum in A549 cells. Furthermore, SanG enhanced the apoptotic effect with doxorubicin in A549 cells. Taken together, our results for the first time provide novel evidence that SanG suppresses proliferation and induces apoptosis via caspase‐3 activation and RPL5 mediated inhibition of c‐Myc with combinational potential with doxorubicin. [ABSTRACT FROM AUTHOR]

Since heat shock protein (HSP27) is a prognostic marker in cervical cancer, in the present study, the apoptotic mechanism of lambertianic acid (LA) was investigated in human cervical cancers in association with HSP27/STAT3/AKT signaling axis. LA exerted significant cytotoxicity, induced sub‐G1 population, and increased the cleavage of Poly (ADP‐ribose) polymerase (PARP) and cysteine aspartyl‐specific protease 3 (caspase3) in HeLa and Caski cancer cells. Consistently, LA downregulated anti‐apopotic genes such as B‐cell lymphoma 2 (Bcl‐2) and inhibitors of apoptosis proteins (c‐IAP) in HeLa and Caski cells. Furthermore, LA‐inhibited phosphorylation of HSP27, signal transducer, and activator of transcription 3 (STAT3) and Protein kinase B (AKT) through disturbing the binding of HSP27 with STAT3 or AKT in HeLa cells. Notably, LA upregulated the level of miR216b in HeLa and Caski cells. Consistently, miR216b mimic suppressed phosphorylation of HSP27 and reduced the expression of pro‐PARP, while miR216b inhibitor reversed the ability of LA to attenuate phosphorylation of AKT, HSP27, and STAT3 and to reduce the expression of pro‐PARP in HeLa cells. Overall, our findings suggest that miRNA216b mediated inhibition of HSP27/STAT3/ AKT signaling axis is critically involved in LA‐induced apoptosis in cervical cancers. [ABSTRACT FROM AUTHOR]

Though honokiol, derived from the Magnolia tree, was known to suppress renal fibrosis, pulmonary fibrosis, non‐alcoholic steatoheptitis, inflammation and cancers, the underlying antifibrotic mechanisms of honokiol are not fully understood in hepatic stellate cells until now. Thus, in the present study, inhibitory mechanism of honokiol on liver fibrosis was elucidated mainly in hepatic stellate cells (HSCs) by 3‐(4, 5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT) assay, cell cycle analysis and western‐blotting. Honokiol exerted cytotoxicity in LX‐2, HSC‐T6 and Hep‐G2 cells. Honokiol increased sub G1 population and activated caspase 3 and cleaved poly (ADP‐ribose) polymerase (PARP) in HSCs. Moreover, honokiol attenuated the expression of alpha smooth muscle actin (α‐SMA), transforming growth factor beta 1 (TGF‐β1), phospho‐Smad3, phospho‐AKT, cyclin D1, c‐Myc, Wnt3a, β‐catenin, and activated phosphorylation of glycogen synthase kinase 3 beta (GSK3β) in HSCs. Conversely, GSK3β inhibitor SB216763 reversed the effect of honokiol on PARP, α‐SMA, phospho‐GSK3β, β‐catenin and sub G1 population in LX‐2 cells. Overall, honokiol exerts apoptotic and antifibrotic effects via activation of GSK3β and inhibition of Wnt3a/β‐catenin signalling pathway. [ABSTRACT FROM AUTHOR]

In the current study, the function of long noncoding RNA (LncRNA) RAB5IF was elucidated in hepatocellular carcinoma (HCCs) in association with LGR5 related signaling. Here TCGA analysis revealed that LncRNA RAB5IF was overexpressed in HCC, and its overexpression level was significantly (p < 0.05) correlated with poor prognosis in patients with HCC. Furthermore, LncRNA RAB5IF depletion suppressed cell proliferation and colony formation, increased sub G1 population, cleavage of poly ADP-ribose polymerase (PARP) and cysteine aspartyl-specific protease (caspase 3) and attenuated the expression of procaspase 3, pro-PARP and B-cell lymphoma 2 (Bcl-2) in HepG2 and Hep3B cells. Furthermore, LncRNA RAB5IF depletion reduced the expression of LGR5 and its downstreams such as β-catenin and c-Myc in HepG2 and Hep3B cells. Notably, LGR5 depletion also attenuated the expression of pro-PARP, pro-caspase3, β-catenin and c-Myc in HepG2 and Hep3B cells. Conversely, LGR5 overexpression upregulated β-catenin and c-Myc in Alpha Mouse Liver 12 (AML-12) normal hepatocytes. Overall, these findings provide novel evidence that LncRNA RAB5IF promotes the growth of hepatocellular carcinoma cells via LGR5 mediated β-catenin and c-Myc signaling as a potent oncogenic target. [ABSTRACT FROM AUTHOR]

Among ribosomal proteins essential for protein synthesis, the functions of ribosomal protein L5 (RPL5) and RPL11 still remain unclear to date. Here, the roles of RPL5 and RPL11 were investigated in association with p53/p21 signaling in the antitumor effect of puromycin mainly in HCT116 and H1299 cancer cells. Cell proliferation assays using 3-[4,5-dimethylthiazole-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assays and colony formation assays, cell cycle analysis, Reverse transcription polymerase chain reaction (RT-PCR) and Western blotting were performed in cancer cells. Puromycin exerted cytotoxic and anti-proliferative effects in p53 wild-type HCT116 more than in p53 null H1299 cells. Consistently, puromycin increased sub-G1, cleaved Poly (ADP-ribose) polymerase (PARP), activated p53, p21, and Mouse double minute 2 homolog (MDM2), and attenuated expression of c-Myc in HCT116 cells. Notably, puromycin upregulated the expression of RPL5 and RPL11 to directly bind to MDM2 in HCT116 cells. Conversely, deletion of RPL5 and RPL11 blocked the activation of p53, p21, and MDM2 in HCT116 cells. Also, puromycin enhanced the antitumor effect with reactivating p53 and inducing tumor apoptosis (RITA) or doxorubicin in HCT116 cells. These findings suggest that puromycin induces p53-dependent apoptosis via upregulation of RPL5 or RPL11 for binding with MDM2, and so can be used more effectively in p53 wild-type cancers by combination with RITA or doxorubicin. [ABSTRACT FROM AUTHOR]

Although shikimic acid from Illicium verum has antioxidant, antibacterial, anti-inflammatory, and analgesic effects, the effect of shikimic acid on lipogenesis has not yet been explored. Thus, in the present study, hypolipogenic mechanism of shikimic acid was examined in HepG2, Huh7 and 3T3-L1 adipocyte cells. Shikimic acid showed weak cytotoxicity in HepG2, Huh7 and 3T3-L1 cells, but suppressed lipid accumulation in HepG2, Huh7 and 3T3-L1 cells by Oil Red O staining. Also, shikimic acid attenuated the mRNA expression of de novo lipogenesis related genes such as FAS, SREBP-1c, and LXR-α in HepG2 cells by RT-PCR analysis and suppressed the protein expression of SREBP-1c and LXR-α in HepG2 and 3T3-L1 cells. It should be noted that shikimic acid activated phosphorylation of AMP-activated protein kinase (AMPK)/Aacetyl-coenzyme A carboxylase (ACC) and reduced the expression of MID1 Interacting Protein 1 (MID1IP1) in HepG2, Huh7 and 3T3-L1 cells. Conversely, depletion of MID1IP1 activated phosphorylation of AMPK, while overexpression of MID1IP1 suppressed phosphorylation of AMPK in HepG2 cells. However, AMPK inhibitor compound c did not affect the expression of MID1IP1, indicating MID1IP1 as an upstream of AMPK. Taken together, our findings suggest that shikimic acid has hypolipogenic effect in HepG2 and 3T3-L1 cells via phosphorylation of AMPK/ACC and inhibition of MID1IP1 as a potent candidate for prevention or treatment of fatty liver and hyperlipidemia. [ABSTRACT FROM AUTHOR]