Apoptosis In KB Cells Research Articles

New Cytotoxic Sesquiterpene Lactones from the Leaves of Tithonia Diversifolia and their Apoptosis Effect Evaluation in KB Cancer Cells.

From the leaves of Tithonia diversifolia, nine sesquiterpenoids (1-9), including two new ones (tagitinin J (1) and tagitinin K (2)) were isolated and structurally determined. Their chemical structures were elucidated by extensive analyses of the HRESIMS and NMR spectral data, as well as comparison with the literature. All of the isolated compounds (except compounds 7-9) significantly exhibited cytotoxic activity against four human cancer cell lines (KB, HepG2, A549 and MCF7), with IC50 values ranging from 0.29-17.0 μM, which were in the same range as the positive control ellipticine or even lower. Further, the apoptosis induction effects of two new compounds 1 and 2 were also investigated and reported. While compound 2 did not induce the apoptosis in KB cells at test concentrations, compound 1 was found to possess anti-proliferative activity through concentration-dependently inducing cell cycle arrest at S phase, morphological changes, activation of caspase 3, and an increase in the early-stage apoptosis of KB cells at a concentration of 7.26 μM.

Kaempferol Inhibits Cervical Cancer Cells by Inducing Apoptosis and Autophagy via Inactivation of the PI3K/AKT/mTOR Signaling Pathway.

Kaempferol, a natural flavonoid, occurs abundantly in fruits and vegetables. It has various bioactivities, with antioxidant, anti-inflammatory, and other beneficial properties. The aim of this study was to investigate the in vitro effects of kaempferol on the proliferation, apoptosis, and autophagy of KB cells, a human cervical cancer cell line, and the corresponding action mechanisms. The inhibitory efficacy of kaempferol on KB cervical cancer cells was investigated through 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, migration assay, 4',6-diamidino-2-phenylindole staining, flow cytometry, acridine orange staining and western blotting. Kaempferol reduced KB cell viability and migration in a dose-dependent manner. Additionally, kaempferol-induced apoptosis was confirmed, and kaempferol treatment influenced levels of apoptotic proteins. Autophagy was detected upon visualization of characteristic autophagic vacuoles and acidic vesicular organelles, and verified using western blotting, which revealed elevated levels of autophagy-related proteins. Kaempferol-mediated apoptosis and autophagy were evidently attributable to reduced phosphorylation in the phosphoinositide 3-kinase (PI3K)/serine/threonine kinase 1 (AKT)/mammalian target of rapamycin (mTOR) pathway. This finding was validated using a pharmacological inhibition assay with the PI3K pathway inhibitor LY294002, which promoted KB cell apoptosis and autophagy. Our results suggest that kaempferol induces apoptosis and autophagy by inhibiting the PI3K/AKT/mTOR pathway in human cervical cancer cells, empirically showing the anticancer effects of kaempferol, and thereby presenting it as a potential anticancer therapeutic agent.

Bioinspired green synthesis of ZnO nanoparticles by marine-derived Streptomyces plicatus and its multifaceted biomedicinal properties

The present study explores the bioinspired green synthesis of zinc oxide nanoparticles (ZnONPs) using marine Streptomyces plicatus and its potent antibacterial, antibiofilm activity against dental caries forming Streptococcus mutans MTCC and S. mutans clinical isolate (CI), cytotoxicity against oral KB cancer cells, hemolysis against blood erythrocytes and artemia toxicity. The bioinspired ZnONPs showed a distinctive absorption peak at 375 nm in UV–Vis spectra, the FT-IR spectra divulged the active functional groups, and XRD confirmed the crystalline nature of the nanoparticles with an average grain size of 41.76 nm. SEM analysis evidenced hexagonal morphology, and EDX spectra affirmed the presence of zinc. The ZnONPs exerted higher antagonistic activity against S. mutans MTCC (Inhibitory zone: 19 mm; MIC: 75 μg/ml) than S. mutans CI (Inhibitory zone: 17 mm; MIC: 100 μg/ml). Results of biofilm inhibitory activity showed a concentration-dependent reduction with S. mutans MTCC (15 %–95 %) more sensitive than S. mutans CI (13 %–89 %). The 50 % biofilm inhibitory concentration (BIC50) of ZnONPs against S. mutans MTCC was considerably lower (71.76 μg/ml) than S. mutans CI (78.13 μg/ml). Confocal Laser Scanning Microscopic visuals clearly implied that ZnONPs effectively distorted the biofilm architecture of both S. mutans MTCC and S. mutans CI. This was further bolstered by a remarkable rise in protein leakage (19 %–85 %; 15 %–77 %) and a fall in exopolysaccharide production (34 mg–7 mg; 49 mg–12 mg). MTT cytotoxicity of ZnONPs recorded an IC50 value of 22.06 μg/ml against KB cells. Acridine orange/ethidium bromide staining showed an increasing incidence of apoptosis in KB cells. Brine shrimp cytotoxicity using Artemia salina larvae recorded an LC50 value of 78.41 μg/ml. Hemolysis assay substantiated the biocompatibility of the ZnONPs. This study underscores the multifaceted application of bioinspired ZnONPs in dentistry.

Marine-derived κ-carrageenan-coated zinc oxide nanoparticles for targeted drug delivery and apoptosis induction in oral cancer.

Kappaphycus alvarezii, a marine red algae species, has gained significant attention in recent years due to its versatile bioactive compounds. Among these, κ-carrageenan (CR), a sulfated polysaccharide, exhibits remarkable antimicrobial properties. This study emphasizes the synergism attained by functionalizing zinc oxide nanoparticles (ZnO NPs) with CR, thereby enhancing its antimicrobial efficacy and target specificity against dental pathogens. In this study, we synthesized ZnO-CR NPs and characterized them using SEM, FTIR, and XRD techniques to authenticate their composition and structural attributes. Moreover, our investigation revealed that ZnO-CR NPs possess better free radical scavenging capabilities, as evidenced by their effective activity in the DPPH and ABTS assay. The antimicrobial properties of ZnO-CR NPs were systematically assessed using a zone of inhibition assay against dental pathogens of S. aureus, S. mutans, E. faecalis, and C. albicans, demonstrating their substantial inhibitory effects at a minimal concentration of 50μg/mL. We elucidated the interaction between CR and the receptors of dental pathogens to further understand their mechanism of action. The ZnO-CR NPs demonstrated a dose-dependent anticancer effect at concentrations of 5μg/mL, 25μg/mL, 50μg/mL, and 100μg/mL on KB cells, a type of Human Oral Epidermal Carcinoma. The mechanism by which ZnO-CA NPs induced apoptosis in KB cells was determined by observing an increase in the expression of the BCL-2, BAX, and P53 genes. Our findings unveil the promising potential of ZnO-CR NPs as a candidate with significant utility in dental applications. The demonstrated biocompatibility, potent antioxidant and antiapoptotic activity, along with impressive antimicrobial efficacy position these NPs as a valuable resource in the ongoing fight against dental pathogens and oral cancer.

Green synthesized zinc oxide nanoparticles from Cinnamomum verum bark extract inhibited cell growth and induced caspase-mediated apoptosis in oral cancer KB cells

IntroductionOral cancer is the sixth most common cancer worldwide with increased mortality cases. Recently, nanomedicines have received greater research attention for cancer treatments. The current exploration focuses to formulate the zinc oxide nanoparticles from C.verum bark (CV-ZnONPs) and assess its in vitro anticancer potentials against oral cancer KB cells. MethodologyThe CV-ZnONPs were synthesized by standard method and then characterized by UV–visible spectrophotometry, XRD, TEM, EDX, FT-IR, and photoluminescence (PL) techniques. The cytotoxicity of CV-ZnONPs against KB cells was assessed using an MTT assay. The level of ROS and MMP in the CV-ZnONPs treated KB cells were measured using DCFH-DA and RH-123 staining, respectively. The dual staining, DAPI, and PI staining were employed to study apoptotic cell death. The comet assay, cell adhesion, and caspase activities were examined, respectively. ResultsOur outcomes confirmed that the CV-ZnONPs have inhibited the KB cell viability. The treatment with the 15 and 20 μg of CV-ZnONPs effectively diminished the MMP and enhanced the ROS status in the KB cells. The outcomes of AO/EB, DAPI, and PI staining confirmed that the formulated CV-ZnONPs triggered apoptosis in the KB cells. The cell adhesion was decreased and caspase activities were improved in the KB cells by the CV-ZnONPs treatment. Discussion and conclusionIn overall, our findings proved that the formulated CV-ZnONPs have shown effective in vitro anticancer activity against oral cancer KB cells, hence it can be a talented anticancer formulation to treat oral cancer in the future.

Inhibitory effect of Solanum xanthocarpum on the growth of KB human oral cancer cell line in vitro through ROS-induced mitochondrial pathway

The aim of the present study was to investigate the effect of Solanum xanthocarpum on KB human oral cancer cells by analyzing its anti-proliferative and apoptotic properties as well as its inhibitory effect on cell adhesion. In this study, the leaves extract of S. xanthocarpum was prepared using the maceration method. Cytotoxic effect of different doses of the S. xanthocarpum extract was assessed using MTT assay. Measurements of reactive oxygen species (ROS), lipid peroxidation and antioxidant enzymes were also performed. In addition, we also studied the impacts of S. xanthocarpum on the apoptosis and mitochondrial membrane potential of KB cells. Determination of antioxidant enzymes and lipid peroxidation was performed using biochemical methods. The S. xanthocarpum showed cytotoxic activity against KB cells with IC50 (200 μg/mL). Besides, DCFH-DA staining and acridine orange/ethidium bromide staining results demonstrated that S. xanthocarpum induced the generation of ROS and apoptosis in KB cells, respectively. Based on the Rh-123 staining results, S. xanthocarpum decreased mitochondrial depolarization in KB cells. Furthermore, the S. xanthocarpum treatment contributed to increased lipid peroxidation, accompanied by reduced activities of superoxide dismutase and catalase, as well as decreased glutathione content. Taken together, these findings indicate that S. xanthocarpum extract might comprise bioactive compounds of therapeutic significance, which can inhibit the growth of KB cells.

The pro-apoptotic and cytotoxic efficacy of polydatin encapsulated poly(lactic-co-glycolic acid) (PLGA) nanoparticles

Polydatin (POL) is an ingredient of many herbal medications and has pharmacological properties. The present study aimed to determine the antitumor potential of Polydatin loaded poly(lactic-co-glycolic acid) (PLGA) nanoparticles (POL-PLGA-NPs) against the cancerous KB cell line. We evaluated the efficiency of synthesized nanoparticles on cell proliferation and apoptosis markers, in addition to the generation of reactive oxygen species (ROS) and its potential to induce damages on the mitochondrial membrane. Morphological changes in the cells (damage to DNA incurred due to cell lysis) were observed under Ethidium Bromide and Acridine orange staining techniques. The results evidenced the loss of cell viability, increased ROS, altered lipid peroxidation levels, increased DNA damage and loss of mitochondrial membrane potential in target cancerous KB cells. Further, immunoblot analysis quantified the expression of caspases-3, -9 and poly(ADP-ribose) polymerase (PARP) also supports the cell damage. Thus, the findings of the study suggested that POL-PLGA-NPs effectively induced oxidative stress by initiating cellular damage leading to apoptosis in KB cells.

Targeted nanoformulation of C1 inhibits the growth of KB spheroids and cancer stem cell-enriched MCF-7 mammospheres

C1, a synthetic analog of curcumin, has been reported to show potent antiproliferative effects against a variety of cancer cells. Here, we report a strong anticancer activity of the folate receptor-targeted lipid nanoparticle formulation of C1 against cancer cells and cancer stem cells both in 2D culture and 3D spheroids. The size of the C1 encapsulated folic acid functionalized nanoliposomes (Lipos-C1) was determined to be 83 ± 17 nm. Lipos-C1 nanoparticles displayed sustained C1 release kinetics at both pH 7.4 and 5.5. The folate receptor (FR) targeted nanoliposomes were internalized into FR-positive KB cells via the folate receptor-mediated endocytosis process. Lipos-C1 killed KB cells much more efficiently than C1. Lipos-C1 depolymerized microtubules, generated ROS, caused DNA damage, and induced apoptosis in KB cells. Importantly, Lipos-C1 strongly inhibited the growth of the 3D KB spheroids than C1. Interestingly, Lipos-C1 also suppressed cancer stem cells (CSCs) enriched MCF-7 mammosphere growth by impeding breast cancer stem cells (BCSCs) enrichment, growth, and proliferation. The results suggested that Lipos-C1 could be a promising nanoformulation for cancer chemotherapy.

Anticancer potential of sclareol against human KB oral carcinoma cell line invitro

Sclareol, a labdane diterpene phytochemical in the various genus Salvia of the herbaceous plant has been anti-carcinogenic properties on numerous human cancer cell lines. Till known the anticancer properties of sclareol on human oral KB cancer cells are not revealed. Therefore, we scrutinized the viability, nucleate damage associated with apoptosis in KB cells treated with sclareol.The cytotoxicity of KB cells displayed to sclareol was evaluated by hexosaminidase and trypan blue exclusion assay. Apoptosis was noticed by Hoechst staining and affirmed by nucleate damage. We discovered that sclareol treatment for KB cells with expressively lessened cell proliferation or viability and evoked cell death in a dose-dependent manner compared with untreated control. Sclareol stimulated inhibition of growing of oral KB cells came along to be induction nucleate damage. Our data proved that sclareol on stimulated apoptosis in human oral KB cancer cells through nucleate damage. These observations recommend the anti-cancer properties of sclareol.

Impaired actin filaments decrease cisplatin sensitivity via dysfunction of volume-sensitive Cl- channels in human epidermoid carcinoma cells.

Cisplatin is a widely used platinum-based anticancer drug in the chemotherapy of numerous human cancers. However, cancer cells acquire resistance to cisplatin. So far, functional loss of volume-sensitive outwardly rectifying (VSOR) Cl- channels has been reported to contribute to cisplatin resistance of cancer cells. Here, we analyzed protein expression patterns of human epidermoid carcinoma KB cells and its cisplatin-resistant KCP-4 cells. Intriguingly, KB cells exhibited higher β-actin expression and clearer actin filaments than KCP-4 cells. The β-actin knockdown in KB cells decreased VSOR Cl- currents and inhibited the regulatory volume decrease (RVD) process after cell swelling. Consistently, KB cells treated with cytochalasin D, which depolymerizes actin filaments, showed smaller VSOR Cl- currents and slower RVD. Cytochalasin D also inhibited cisplatin-triggered apoptosis in KB cells. These results suggest that the disruption of actin filaments cause the dysfunction of VSOR Cl- channels, which elicits resistance to cisplatin in human epidermoid carcinoma cells.

β-Caryophyllene promotes oxidative stress and apoptosis in KB cells through activation of mitochondrial-mediated pathway – An in-vitro and in-silico study

Beta-caryophyllene (BCP), are natural bicyclic sesquiterpenes which are present in numerous plants worldwide. BCP has antioxidant, antimicrobial, and antifungal properties. Here, we studied its anticancer, anti-inflammatory, and cytotoxic effects. Cells treated with BCP, in a dose-dependent manner, exhibited morphological changes, showed lower cell growth, underwent apoptosis and lost the ability to metastasis through the suppression of NF-ҡ B via PI3K/AKT signalling pathway. These results elucidate that the inhibition of NF-ҡ B and PI3K/AKT is one of the most important mechanism by which BCP suppresses cancer cell proliferation and enhances apoptosis.

Porphyromonas gingivalis degrades integrin β1 and induces AIF-mediated apoptosis of epithelial cells

Background: Porphyromonas gingivalis, a major pathogen of chronic periodontitis, adheres to and invades epithelial cells via an interaction between fimbriae and integrin. P. gingivalis proliferation and infection may affect the survival of cells. In this study, we further examined alternative signaling pathways mediating epithelial-cell death induced by P. gingivalis and the role of the cell-adhesion molecule integrin.Methods: Human epithelial KB cells interacted with P. gingivalis to evaluate cell death by Annexin V-propidium iodide (PI) staining. JC-1 staining was used to measure mitochondrial membrane potential (MMP). The mRNA and protein of integrin β1, apoptosis-inducing factor (AIF) and caspase-3 were detected by real-time PCR and western blot. Caspase-3 activity was analyzed by spectrophotometry.Results: P. gingivalis infection downregulated integrin β1 and led to cell detachment in a dose and time-dependent manner. Large amount of P. gingivalis induced MMP depolarization and apoptosis in KB cells. Moreover, P. gingivalis up-regulated AIF, but not activate caspase-3 during apoptosis. In addition, AIF inhibitor N-Phenylmaleimide almost inhibited the P. gingivalis-induced apoptosis.Conclusions: P. gingivalis disrupts epithelial-cell adhesion by degrading integrin β1 and induces caspase-independent, AIF-mediated mitochondrial apoptosis, which may promote the damage of oral tissue.

Effects of chrysin on the apoptosis in oral squamous carcinoma KB cell line and the underlying mechanisms

To investigate the effect of chrysin on apoptosis of oral squamous carcinoma KB cell line and the possible mechanisms, and to provide new ideas for the treatment of oral cancer. Methods: Oral cancer KB cells were treated with different concentrations of chrysin (1, 2, 4, 8, 16, and 32 μmol/L) for 24 h. Cell proliferation was detected by MMT assay; apoptosis was detected by flow cytometry; the activity of caspase-3/7 was detected by chemiluminescent assay; mitochondrial membrane potential in KB cells was determined by JC-1 assay; and Western blotting was used to determine the activation of protein kinase B (AKT) and phosphoinositide-3-kinase (PI3K). Results: Chrysin inhibited the proliferation of KB cells in a concentration-dependent manner, accompanied by increase in apoptosis of KB cells, activation of caspase-3/7, decrease in mitochondrial membrane potential, and suppression of the phosphorylation of AKT and PI3K. Conclusion: The effect of chrysin on KB cell apoptosis may be related to mitochondrial dysfunction and inhibition of PI3K/AKT pathway.

Design and characterization of a novel Arthrospira platensis glutathione oxido-reductase-derived antioxidant peptide GM15 and its potent anti-cancer activity via caspase-9 mediated apoptosis in oral cancer cells

Glutathione oxido-reductase (GR) is a primary antioxidant enzyme of most living forms which protects the cells from oxidative metabolism by reducing glutathione (GSH) from its oxidized form (GSSG). Although the antioxidant role of the enzyme is well characterized, the specific role of conserved N' peptide sequence in antioxidant mechanism remains unclear. In this study, we have identified an RNA sequence encoding GR enzyme from spirulina, Arthrospira platensis (Ap) and the changes in its gene expression profile was analysed during H2O2 stress. Results showed that H2O2 (10 mM) stimulated the expression of ApGR throughout the timeline of study (0, 5, 10, 15 and 20 days) with highest expression at 5th day post-exposure which confirmed the antioxidant role of ApGR in spirulina during H2O2 induced oxidative stress. A dithiol containing short antioxidant peptide, 39GGTCVIRGCVPKKLM53 (GM15) from ApGR was predicted and its radicals (superoxide and hydroxyl radical) scavenging potential was confirmed by in vitro cell-free assays. GM15 (12.5 μM) reduced the intracellular generalized oxidative stress level, as measured using DCFDA assay in H2O2 exposed leucocytes without affecting any of the cellular population. Further, the biomedical application of the radical scavenging property of GM15 was validated in oral carcinoma (KB) cells where GM15 exhibited significant cytotoxicity. Also, GM15 exhibited heterogenous effects on intracellular oxidative stress level in KB cells: at lower concentration (6.25 μM), the peptide reduced oxidative stress whereas, at higher concentration (25 μM) it increased the intensity of oxidative stress. GM15 (25 μM) induced caspase-9 mediated apoptosis in KB cells along with membrane disruption and DNA degradation which are confirmed by propidium iodide (PI) internalization and comet assays, respectively. Overall, the study shows that GM15 peptide i) scavenges superoxide, hydroxyl radicals, and influences intracellular oxidative stress, and ii) has anti-cancer effect in oral cancer cells.

Grail suppresses stress-induced apoptosis and colony formation in oral cancer cells

Aims: The purpose of this study is to investigate the role of Grail in stress-induced apoptosis and tumorigenesis in oral cancer cells. Subjects and Methods: We analyzed gene expression of Grail in oral cancer cell lines (KB, SAS, SCC-4, and SCC-25) treated with 5-azadC or/and trichostatin A (TSA) by RT-PCR. The effects of Grail on the expression of p53 and p21 were analyzed by real-time polymerase chain reaction and Western blot for KB cells (KB/vector, KB/Grail and KB/shGrail). The anti-apoptotic effects of Grail were determined by fluorescence-activated cell sorting in KB/vector and KB/Grail cells. The effects of Grail on tumorigenesis were through clonogenic analysis in KB cells (KB/vector, KB/Grail and KB/shGrail). Results: Treatment with 5-azadC or/and TSA-induced Grail mRNA expression in oral cancer cells. Grail overexpression reduced p53 and p21 expression. Conversely, p53 and p21 expressions were increased in KB/shGrail cells. Furthermore, Grail can inhibit resveratrol-or etoposide-induced apoptosis in KB cells. Finally, Grail can significantly suppress colony formation in KB cells. Conclusions: The expression of Grail is epigenetically regulated in oral cancer cells. Grail can reduce p53 and p21 expression and stress-induced cell death and suppress the colony formation in KB cells.

Cytotoxic Effects of Pistacia Atlantica (Baneh) Fruit Extract on Human KB Cancer Cell Line.

Plants with anticancer properties are considered as cancer preventive and treatment sources, due to their some biological effects. Apoptosis induction and anti-proliferative effects of Baneh extract on various cancer cell lines have been reported. Hence, this study was designed to evaluate the cytotoxic effects of this fruit on KB and human gingival fibroblast cell lines (HGF). KB and HGF cells were treated with various concentrations of ethanolic Baneh extract and cisplatin as positive control. Cytotoxic activity and apoptosis induction were investigated using WST-1 and Annexin V assays. Data were analyzed using ANOVA and student's t-tests. IC50 after 24 and 48 hours treatment were respectively 2.6 and 1 mg/mL for KB cell line, and 1.5 and 1.6 mg/mL for HGF cell. During 48 hours Baneh extract induced apoptosis without significant necrosis, in a time- and dose-dependent manner. The induction of apoptosis in KB cells was significantly higher than HGF. It seems that ethanolic extract of Baneh contains compounds that can suppress KB cell growth through the induction of apoptosis. Within 48 hours, less cytotoxic effects were observed on normal fibroblast cells; therefore, it might be a potential anticancer agent.

Abrin, a Type II Ribosome Inactivating Protein: Differential Cytotoxicity and Development of the Vaccine Against its Lethality

The plant toxin, Abrin, a type II ribosome inactivating protein, is extremely lethal, the human fatal dose being ~1 μg/kg body weight. Abrin has been classified as an agent for bioterrorism, which is of huge concern. On the other hand, the high toxic property of abrin has been employed in generating immunotoxins, where its toxin moiety is conjugated to cell surface marker specific antibodies for cell targeted killing.Different cell types exhibit variable levels of sensitivity to abrin toxicity. To gain insights into this, two cell lines requiring strikingly different IC50 of abrin for inactivating ribosomes were studied. In KB cells, the abrin A chain was found to be sequestered from the cytoplasm into the nucleus by a protein called Brain acid soluble protein (BASP‐1), enabling the cells to resist abrin‐mediated inhibition of protein synthesis. In Ovcar‐3 cells that express negligible amounts of BASP‐1, the A chain remains in the cytosol to exert its N‐glycosidase activity, therefore succumb to low concentrations of abrin. In contrast, when the apoptosis inducing activity of abrin was examined, both the cells showed comparable apoptotic population. Employing conjugates of the wild type and active site mutant ABA with the ricin B chain it was found that abrin‐induced apoptosis was dependent on inhibition of protein synthesis leading to ER‐stress in Ovcar‐3 cells, but not in KB cells. Direct DNA damage caused by the abrin A chain sequestered in the nucleus might be contributing to the apoptosis in KB cells.Towards establishing vaccines to counteract abrin‐induced lethality, we designed a recombinant chimeric protein of Abrin A chain and Abrus precatorius agglutinin A chain on the basis of epitope specificity. Abrin part of the chimera contains the epitope for the only two neutralizing antibodies; D6F10 and A7C4, reported against abrin till now. Importantly, the chimera harboured no enzymatic activity and was found to protect 100 % mice from abrin challenge upto 45 X LD50 dose. Thus the chimera appears to be potential vaccine against abrin induced lethality.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Retracted: Tanshinone IIA Induces Apoptosis in Human Oral Cancer KB Cells through a Mitochondria-Dependent Pathway.

[This retracts the article DOI: 10.1155/2014/540516.].

Folate acid-Cyclodextrin/Docetaxel induces apoptosis in KB cells via the intrinsic mitochondrial pathway and displays antitumor activity in vivo

Docetaxel (DTX) is an antitumor therapeutic drug limited by solubility and selective delivery tissues. We previously prepared DTX/folate acid-Cyclodextrin (FA-CD) inclusion complexes that target folate receptors of tumor cells and showed that these complexes inhibited cancer cell proliferation by inducing apoptosis. Here we further determined the antitumor effect and apoptotic mechanism of DTX/FA-CD. DTX/FA-CD induced reactive oxygen species-mediated mitochondrial apoptosis in KB cells. DTX/FA-CD led to apoptosis accompanied with the repression of mitochondrial membrane potential and glutathione and overexpression of reactive oxygen species and catalase. DTX/FA-CD also specifically accumulated into tumor regions in KB tumor-bearing mice by active targeting. DTX/FA-CD significantly suppressed tumor growth and showed low toxicity in KB tumor xenografts. We concluded that the DTX/FA-DTX inclusion complex induced the intrinsic mitochondrial-mediated apoptosis to cause cell death. Our results showed favorable antitumor effects of DTX/FA-DTX and indicate DTX/FA-DTX could serve as a safe and effective delivery system for antitumor therapy.

Umbelliferone arrest cell cycle at G0/G1 phase and induces apoptosis in human oral carcinoma (KB) cells possibly via oxidative DNA damage

Umbelliferone (UMB) has widespread pharmacological activity, comprising anti-inflammatory, anti-oxidant, anti-genotoxic and anti-immunomodulatory but the anticancer activity remains unknown in human oral carcinoma (HOC) KB cells. MTT assay determinations was revealed that treatment of KB cells with UMB, prevent and reduce the cell proliferation with the IC50 − 200μM as well as induces loss of cell viability, morphology change and internucleosomal DNA fragmentation in a concentration dependent manner. Acridine orange and ethidium bromide dual staining assay established that UMB induced apoptosis in KB cells in a dose dependent manner. Alkaline comet assay determination revealed UMB has the potential to increase oxidative DNA damage in KB cells through DNA tail formation significantly (p<0.05). Furthermore, UMB brought a dose-dependent elevation of reactive oxygen species (ROS), which is evidenced by the DCF fluorescence, altered the mitochondrial membrane potential in KB cells. Similarly, we observed increased DNA damage stimulated apoptotic morphological changes in UMB treated cells. Taken together, the present study suggests that UMB exhibits anticancer effect on KB cell line with the increased generation of intracellular ROS, triggered oxidative stress mediated depolarization of mitochondria, which contributes cell death via DNA damage as well as cell cycle arrest at G0/G1 phase. The results have also provided us insight in the pharmacological backgrounds for the potential use of UMB, to target divergent pathways of cell survival and cell death. To conclude UMB could develop as a novel candidate for cancer chemoprevention and therapy, which is our future focus and to develop a connectivity map between in vivo and in vitro activity.