SrfABC Toxin from Xenorhabdus stockiae Induces Cytotoxicity and Apoptosis in HeLa Cells

Our previous study showed that the srfABC operon, which was originally identified in Salmonella enterica as an SsrB-regulated operon clustered with the flagellar class 2 operon, exhibited significant cytotoxicity against insect midgut CF-203 cells and injectable insecticidal activity against Helicoverpa armigera larvae. The srfABC operon was widely distributed among bacteria, which raises the question of their biological roles in different species. In this study, we investigated the cytotoxic effect of SrfABC toxin on mammalian cell lines. When simultaneously expressed in the Escherichia coli cytoplasm, SrfABC exhibited cytotoxicity against all tested mammalian cancer cell lines (B16, 4T-1, Hep-3B, and HeLa) in a dose-dependent manner. Intracellular expression of SrfA–FLAG, SrfB–FLAG, or SrfC–FLAG also resulted in inhibition of proliferation and apoptosis on HeLa cells. When incubated with HeLa cells separately, SrfA, SrfB, and SrfC proteins alone could enter HeLa cells, then induce apoptosis and cytotoxicity. SrfC protein shifts its localization from cytoplasm to nucleus with the aid of SrfA and/or SrfB protein. Although SrfA, SrfB, and SrfC proteins alone exhibited a cytotoxic effect against HeLa cells, all three components were essential for the full cytotoxicity. Native PAGE and co-immunoprecipitation assay demonstrated that SrfA, SrfB, and SrfC proteins could interact with each other and form a heteromeric complex.

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Cytotoxic effect of Montivipera bornmuelleri’s venom on cancer cell lines: in vitro and in vivo studies

PeerJ ◽

10.7717/peerj.9909 ◽

2020 ◽

Vol 8 ◽

pp. e9909

Author(s):

Carol Haddoub ◽

Mohamad Rima ◽

Sandrine Heurtebise ◽

Myriam Lawand ◽

Dania Jundi ◽

...

Keyword(s):

Tumor Growth ◽

Cell Lines ◽

Cytotoxic Effect ◽

In Vivo Studies ◽

Dependent Manner ◽

In Vivo Testing ◽

Murine Fibrosarcoma ◽

Dose Dependent

Background Montivipera bornmuelleri’s venom has shown immunomodulation of cytokines release in mice and selective cytotoxicity on cancer cells in a dose-dependent manner, highlighting an anticancer potential. Here, we extend these findings by elucidating the sensitivity of murine B16 skin melanoma and 3-MCA-induced murine fibrosarcoma cell lines to M. bornmuelleri’s venom and its effect on tumor growth in vivo. Methods The toxicity of the venom on B16 and MCA cells was assessed using flow cytometry and xCELLigence assays. For in vivo testing, tumor growth was followed in mice after intratumoral venom injection. Results The venom toxicity showed a dose-dependent cell death on both B16 and MCA cells. Interestingly, overexpression of ovalbumin increased the sensitivity of the cells to the venom. However, the venom was not able to eradicate induced-tumor growth when injected at 100 µg/kg. Our study demonstrates a cytotoxic effect of M. bornmuelleri’s venom in vitro which, however, does not translate to an anticancer action in vivo.

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Cytotoxic Effect on Cancer Cells and Structural Identification of Phenols from Spatholobi Caulis by HPLC-ESI-MSn

Natural Product Communications ◽

10.1177/1934578x0900400614 ◽

2009 ◽

Vol 4 (6) ◽

pp. 1934578X0900400 ◽

Cited By ~ 1

Author(s):

Dan Lu ◽

Hua He ◽

Bin Wu ◽

Shanjing Yao

Keyword(s):

Cancer Cells ◽

Cell Lines ◽

Cancer Cell ◽

Cytotoxic Effect ◽

Human Cancer ◽

Cancer Cell Lines ◽

Screening Tests ◽

Hl60 Cell ◽

Dependent Manner ◽

Dose Dependent

To find fractions from Spatholobi caulis with cytotoxic effects on cancer cell lines, screening tests were carried out using the SRB assay on the HL60 cell line. Further investigation with HL60 and another four human cancer cell lines (KB, K562, MCF-7 and Hep G2), revealed a dose-dependent response. High-performance liquid chromatography coupled with electrospray ionization-tandem mass spectrometry (HPLC- ESI-MSn) was used to isolate and identify the active constituents from the active fraction. Three fractions (F-IV, F-V and F-VII) showed in vitro cytotoxicity. F-V inhibited the growth of cancer cells in a dose-dependent manner. The IC50 values for KB, K562 and HL60 cells were 17.6, 8.3 and 9.7 μg/mL, respectively. The dominating constituents of F-V were either identified or tentatively characterized as nine phenolic compounds, eight isoflavones and 9-methoxycoumestrol. The isoflavones 7-hydroxy-3′,4′-dimethoxy isoflavone, 7-hydroxy-6,2′,4′-trimethoxy isoflavone and 3′-hydroxy-7,4′-dimethoxy isoflavone are reported for the first time for Spatholobi caulis. The results suggest that these compounds contribute to the cytotoxic effect of Spatholobi caulis.

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Proteosome Inhibitor Treatment Down-Regulates S-Phase Kinase-Associated Protein 2 Causing Inhibition of Proliferation and Induces Apoptosis in Primary Effusion Lymphoma.

Blood ◽

10.1182/blood.v108.11.4616.4616 ◽

2006 ◽

Vol 108 (11) ◽

pp. 4616-4616

Author(s):

Azhar R. Hussain ◽

Naif A. Al-Jomah ◽

Mehar Sultana ◽

Manugaran S. Pulicat ◽

Khawla S. Al-Kuraya ◽

...

Keyword(s):

Cell Cycle ◽

Cell Lines ◽

Primary Effusion Lymphoma ◽

S Phase ◽

Dependent Manner ◽

Down Regulation ◽

Inhibition Of Proliferation ◽

Dose Dependent ◽

Inhibitor Treatment

Abstract Proteosome inhibition is a novel approach for treating malignancy and has been approved for clinical use. The proteosome is the primary proteolytic mechanism in eukaryotic cells and inhibition of its catalytic activity initiates a cascade of events affecting cell cycle and apoptotic activities. These activities ultimately lead to cell cycle arrest and apoptosis in malignant cells however, the normal counterpart of these cells are spared. In this study, we used a panel of primary effusion lymphoma cell lines (BC1, BC3, BCBL1 and HBL6) to study the effects of proteosome inhibitor, MG132 on cell proliferation and apoptosis. Our data showed that proteosome inhibitor MG132 decreased cell viability as well as induced apoptosis in a dose dependent manner ranging from 0.5–10μM. Furthermore, treatment with 2.5μM MG132 for 24hours induced 41% apoptosis in BC1, 51% in BC3, 41% in BCBL1 and 48% in HBL6 cell lines as detected by annexinV/PI dual staining. S-phase kinase-associated protein 2 (skp-2) is a proto-oncogene and over expressed in various types of tumors. We sought to determine the role of Skp-2 following proteosome inhibition in PELs. MG132 treatment of PEL cell lines resulted in down-regulation of SKP-2 protein in a dose dependent manner whereas the expression of p-27 was up-regulated demonstrating an inverse relationship between these two proteins. Furthermore, MG132 treatment of PELs led to conformational changes in Bax protein and translocation to the mitochondria leading to the loss of mitochondrial membrane potential with subsequent release of cytochrome c from mitochondria into cytosol. Cytochrome c release caused activation of caspase-3 followed by polyadenosin-5′-diphosphate-ribose polymerase (PARP) cleavage. In addition, proteosome inhibitor treatment also caused down-regulation of inhibitor of apoptosis protein, XIAP. Taken together, our findings show that proteosome inhibition causes down-regulation of skp-2, up-regulation of p-27, inhibition of proliferation as well as caspase-dependent apoptosis in primary effusion lymphoma cells suggesting a role of proteosome inhibitors in the treatment of these aggressive cancers.

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Antheraea proylei J. sericin induces apoptosis in a caspase dependent manner in A549 and HeLa cells and caspase independent manner in PC3 cells

10.21203/rs.3.rs-56346/v2 ◽

2021 ◽

Author(s):

Potsangbam Jolly Devi ◽

Asem Robinson Singh ◽

Lisam Shanjukumar Singh ◽

Laishram Rupachandra Singh ◽

Sanjenbam Kunjeshwori Devi

Keyword(s):

Cell Cycle ◽

Cell Lines ◽

Hela Cells ◽

Molecular Mechanisms ◽

Human Lung Cancer ◽

Treatment Modalities ◽

Dependent Manner ◽

Pc3 Cells ◽

Mulberry Silkworm ◽

Dose Dependent

Abstract BackgroundIn spite of much progress in understanding the biology of cancer disease, advancement in technology for early diagnosis, the expanding array of anticancer drugs and treatment modalities, the global cancer burden is still significant and increasing. It is estimated that the new cases of cancer in the year 2040 will be 29.4 million per year globally. Sericin, an adhesive protein of silk cocoons has been shown to be a potential protein in various biomedical applications including cancer therapeutics. The present study evaluates the anticancer property of sericin from cocoons of Antheraea proylei J. (SAP) against human lung cancer (A549), cervical cancer (HeLa), and prostate cancer (PC3) cell lines. This is the first report of anti-cancer activity of the non-mulberry silkworm A. proylei J. Methods SAP was prepared from cocoons of A. proylei J. by the process of degumming method. The amino acid composition of SAP was determined by HPLC. Cytotoxicity activity was assessed by MTT assay and genotoxicity activity was assessed by comet assay. Cleavage of caspase and PARP proteins and phosphorylation of MAPK pathway members were analysed by Western blotting. Cell cycle analysis was done by flow cytometery.ResultsSAP causes cytotoxicity to A549, HeLa and PC3 cell lines with the IC50 values ranging from 3.4-3.9 µg/µl. SAP induces apoptosis in a dose-dependent manner through caspase-3 and p38 and ERK pathways in A549 and HeLa cells respectively whereas in PC3 cells, SAP induces apoptosis independent of caspase through p38 pathway. Moreover, in the case of A549 and HeLa cells SAP induces cell cycle arrest at S phase in a dose dependent manner whereas at G0 phase in the case PC3 cells.ConclusionSAP induces apoptosis in A549, HeLa, and PC3. The difference in the molecular mechanisms of apoptosis induced by SAP in A549 and HeLa and in PC3 may be due to the differences in the genotypes of the cancer cell lines. However, further investigation is warranted. The overall results of the present study envisage the possibility of using SAP as anti-tumorigenic agent.

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Sericin Preparation from Cocoons of Oak Tasar Silkworm Antheraea proylei J. Induces Apoptosis in a Caspase-dependent Manner in A549 and HeLa Cells and Caspase-independent Manner in PC3 Cells

10.21203/rs.3.rs-56346/v1 ◽

2020 ◽

Author(s):

Potsangbam Jolly Devi ◽

Asem Robinson Singh ◽

Lisam Shanjukumar Singh ◽

Laishram Rupachandra Singh ◽

Sanjenbam Kunjeshwori Devi

Keyword(s):

Cell Cycle ◽

Cell Lines ◽

Hela Cells ◽

Human Lung Cancer ◽

Treatment Modalities ◽

Dependent Manner ◽

Hela Cell Lines ◽

Pc3 Cells ◽

Mulberry Silkworm ◽

Dose Dependent

Abstract BackgroundDespite much progress in understanding the biology of cancer disease, advancement in technology for early diagnosis, the expanding array of anticancer drugs, and treatment modalities, the global cancer burden is still significant and increasing. It is estimated that the new cases of cancer in the year 2040 will be 29.4 million per year globally. Sericin, an adhesive protein of silk cocoon, is a potential protein in various biomedical applications including cancer therapeutics. The present study evaluates the anticancer property of sericin prepared from cocoons of Antheraea proylei J. (A. proylei ) against human lung cancer (A549), cervical cancer (HeLa), and prostate cancer (PC3) cell lines. This is the first report of the anti-cancer activity of the non-mulberry silkworm A. proylei. MethodsSericin preparation (SP) was prepared from cocoons of A. proylei J. by the process of the degumming method. The amino acid composition of the SP was determined by HPLC. Cytotoxicity activity was assessed by MTT assay and genotoxicity activity was assessed by comet assay. Cleavage of caspase and PARP proteins and phosphorylation of MAPK pathway members were analyzed by Western blotting. Cell cycle analysis was done by FACS flow cytometry.Results SP causes cytotoxicity to A549, HeLa, and PC3 cell lines with the IC50 values ranging from 3.4-3.9 µg/µl. SP induces apoptosis in a dose-dependent manner through caspase-3 and p38/SAPK/ERK pathways in A549 and HeLa cells whereas in PC3 cells SP induces apoptosis independent of caspase but through p38 pathway. Moreover, in the case of A549 and HeLa cells, SP induces cell cycle arrest at the S phase whereas at the G0 phase in the case of PC3 cells in a dose-dependent manner.ConclusionThe difference in the molecular mechanisms of apoptosis induced by SP in A549 and HeLa cell lines, and in PC3 cell lines may be due to the difference in the genotypes of the cancer cell lines where A549 and HeLa cells are being non-malignant and p53 positive whereas PC3 cell is being malignant and p53 negative. The overall results of the present study envisage the possibility of using SP as an anti-tumorogenic agent.

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Inhibition of p38 Mitogen-Activated Protein Kinase Impairs Mayaro Virus Replication in Human Dermal Fibroblasts and HeLa Cells

Viruses ◽

10.3390/v13061156 ◽

2021 ◽

Vol 13 (6) ◽

pp. 1156

Author(s):

Madelaine Sugasti-Salazar ◽

Yessica Y. Llamas-González ◽

Dalkiria Campos ◽

José González-Santamaría

Keyword(s):

Protein Expression ◽

Hela Cells ◽

Plaque Assay ◽

Dermal Fibroblasts ◽

Dependent Manner ◽

Human Dermal Fibroblasts ◽

Mitogen Activated Protein ◽

Mayaro Virus ◽

The Impact ◽

Dose Dependent

Mayaro virus (MAYV) hijacks the host’s cell machinery to effectively replicate. The mitogen-activated protein kinases (MAPKs) p38, JNK, and ERK1/2 have emerged as crucial cellular factors implicated in different stages of the viral cycle. However, whether MAYV uses these MAPKs to competently replicate has not yet been determined. The aim of this study was to evaluate the impact of MAPK inhibition on MAYV replication using primary human dermal fibroblasts (HDFs) and HeLa cells. Viral yields in supernatants from MAYV-infected cells treated or untreated with inhibitors SB203580, SP600125, U0126, or Losmapimod were quantified using plaque assay. Additionally, viral protein expression was analyzed using immunoblot and immunofluorescence. Knockdown of p38⍺/p38β isoforms was performed in HDFs using the PROTACs molecule NR-7h. Our data demonstrated that HDFs are highly susceptible to MAYV infection. SB203580, a p38 inhibitor, reduced MAYV replication in a dose-dependent manner in both HDFs and HeLa cells. Additionally, SB203580 significantly decreased viral E1 protein expression. Similarly, knockdown or inhibition of p38⍺/p38β isoforms with NR-7h or Losmapimod, respectively, affected MAYV replication in a dose-dependent manner. Collectively, these findings suggest that p38 could play an important role in MAYV replication and could serve as a therapeutic target to control MAYV infection.

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Baicalein Inhibits the Proliferation of Cervical Cancer Cells Through the GSK3β-Dependent Pathway

Oncology Research Featuring Preclinical and Clinical Cancer Therapeutics ◽

10.3727/096504017x15031557924141 ◽

2018 ◽

Vol 26 (4) ◽

pp. 645-653 ◽

Cited By ~ 4

Author(s):

Xiaoling Wu ◽

Zhiqin Yang ◽

Huimin Dang ◽

Huixia Peng ◽

Zhijun Dai

Keyword(s):

Cell Cycle ◽

Cervical Cancer ◽

Cyclin D1 ◽

Cancer Cells ◽

Hela Cells ◽

Glycogen Synthase ◽

Dependent Manner ◽

Cervical Cancer Cells ◽

Dose Dependent ◽

Dependent Pathway

Baicalein, a flavonoid derived from the root of Scutellaria baicalensis, has been reported to possess multiple pharmacological activities, such as anticancer and anti-inflammatory properties. This study investigated the effect of baicalein in cervical cancer cells. Cell growth curve and MTT assay were performed and revealed that baicalein inhibited the proliferation of SiHa and HeLa cells in a dose-dependent manner. We further found that baicalein arrested the cell cycle of SiHa and HeLa cells at the G0/G1 phase by suppressing the expression of cyclin D1 through the downregulation of phosphorylated protein kinase B (p-AKT) and phosphorylated glycogen synthase kinase 3β (p-GSK3β) according to FACS assays and Western blotting. Moreover, when CHIR-99021, a GSK3β inhibitor, was added to baicalein-treated SiHa cells, the expression of cyclin D1 was recovered, and cell proliferation was promoted. In conclusion, these data indicated that baicalein suspended the cell cycle at the G0/G1 phase via the downregulation of cyclin D1 through the AKT‐GSK3β signaling pathway and further inhibited the proliferation of SiHa and HeLa cervical cancer cells.

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SUN-132 KLF5 Is a Poor Prognostic Marker and Therapeutic Target for Middle Eastern Papillary Thyroid Carcinoma

Journal of the Endocrine Society ◽

10.1210/jendso/bvaa046.1213 ◽

2020 ◽

Vol 4 (Supplement_1) ◽

Author(s):

Khawla S Al-Kuraya ◽

Abdul K Siraj ◽

Pratheeshkumar Poyil ◽

Divya Padmaja ◽

Sandeep Kumar Parvathareddy ◽

...

Keyword(s):

Thyroid Cancer ◽

Papillary Thyroid Carcinoma ◽

Thyroid Carcinoma ◽

Cell Lines ◽

Therapeutic Target ◽

Critical Role ◽

Dependent Manner ◽

Papillary Thyroid ◽

Inhibition Of Proliferation ◽

Over Expression

Abstract Thyroid cancer is the second most common malignancy among females in Saudi Arabia, with Papillary thyroid carcinoma (PTC) accounting for 80-90%. The Kruppel-like factor 5 (Klf5) is a transcription factor that play a critical role in cell transformation, proliferation and oncogenesis. Immunohistochemical analysis of KLF5 was performed in 1219 PTC cases. KLF5 over-expression was noted in 65.1% (793/1219) of PTCs, and was significantly associated with tall-cell variant (p <0.0001), extrathyroidal extension (p = 0.0003), lymph node metastasis (p < 0.0001) and stage IV tumors (p < 0.0001). Significant association was also noted with HIF-1α over-expression (p = 0.0492). Interestingly, KLF5 over-expressing tumors showed poor disease-free survival (p = 0.0066). Functional studies in PTC cell lines showed that KLF5 co-immunoprecipitated with HIF-1α. Knockdown of KLF5 decreased the expression of HIF-1α while KLF5 was not affected by HIF-1α inhibition, suggesting that KLF5 is a functional upstream of HIF-1α. Down-regulation of KLF5 using specific inhibitor, ML264 or siRNA inhibited cell invasion and migration. In addition, treatment of PTC cell lines with ML264 resulted in inhibition of proliferation and induction of apoptosis in a dose-dependent manner. Furthermore, silencing of KLF5 significantly decreased the self-renewal ability of spheroids generated from PTC cells. Our findings confer that KLF5 may be a potential therapeutic target for the treatment of papillary thyroid cancer.

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In vitro cytotoxic study for partially purified resveratrol extracted from grape skin fruit Vitis vinifera

Journal of Biotechnology Research Center ◽

10.24126/jobrc.2009.3.2.66 ◽

2009 ◽

Vol 3 (2) ◽

pp. 40-47

Author(s):

Zainab Y. Mohammed ◽

Essam F. Al-Jumaily ◽

Nahi Y. Yaseen

Keyword(s):

Cell Line ◽

Cell Lines ◽

Cytotoxic Effect ◽

Inhibitory Effect ◽

Mammary Adenocarcinoma ◽

Dependent Manner ◽

Grape Skin ◽

Grape Fruit ◽

Glass Column

The partial purified resveratrol was obtained from the skin of black grape fruit cultivated in Iraq using 80% ethanolic solution, then an acid hydrolysis with 10% HCl solution for (10–30) min at 60Cº was carried out. The aglycone moiety was taken with an organic solvent (chloroform), then using an open glass column packed with silica gelG 60 as a stationary phase and a mobile phase of; benzene: methanol: actic acid (20:4:1). The study utilized an in vitro evaluation for the cytotoxic effect of the partially purified resveratrol on some cell lines including, the murine mammary adenocarcinoma (Ahmed –Mohammed –Nahi–2003 -AMN -3) cell line; the human laryngeal carcinoma (Hep -2) cell line and the Rat Embryo Fibroblast (REF) cell line at different concentrations and different exposure time of treatment. The partial purified resveratrol extract concentrations ranging (7.8–4000) µg/ml in a two fold serial dilutions were used to treat the three types of cell lines for 48 and 72 hours intervals. AMN-3 cell lines showed highest sensitivity toward the cytotoxic effect of the paritial purified resveratrol than other cell lines after 48 hours in a dose dependent manner. While Hep-2 cell line showed novel behavior, the lowest concentration of cell treatment gave the most significant (P< 0.01) inhibitory effect. Only the highest concentration gave significant inhibitory effect (P< 0.01) with the transformed Ref cell line.

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NPM1 is a Novel Therapeutic Target and Prognostic Biomarker for Ewing Sarcoma

Frontiers in Genetics ◽

10.3389/fgene.2021.771253 ◽

2021 ◽

Vol 12 ◽

Author(s):

Yangfan Zhou ◽

Yuan Fang ◽

Junjie Zhou ◽

Yulian Liu ◽

Shusheng Wu ◽

...

Keyword(s):

Ewing Sarcoma ◽

Es Cells ◽

Differentially Expressed Gene ◽

Bioinformatic Analysis ◽

Dependent Manner ◽

Hub Gene ◽

Immune Infiltration ◽

Proliferation And Apoptosis ◽

Immune Related Genes ◽

Dose Dependent

Ewing sarcoma (ES) is a cancer that may originate from stem mesenchymal or neural crest cells and is highly prevalent in children and adolescents. In recent years, targeted therapies against immune-related genes have shown good efficacy in a variety of cancers. However, effective targets for immunotherapy in ES are yet to be developed. In our study, we first identified the immune-associated differential hub gene NPM1 by bioinformatics methods as a differentially expressed gene, and then validated it using real time-PCR and western blotting, and found that this gene is not only closely related to the immune infiltration in ES, but also can affect the proliferation and apoptosis of ES cells, and is closely related to the survival of patients. The results of our bioinformatic analysis showed that NPM1 can be a hub gene in ES and an immunotherapeutic target to reactivate immune infiltration in patients with ES. In addition, treatment with NPM1 promoted apoptosis and inhibited the proliferation of ES cells. The NPM1 inhibitor NSC348884 can induce apoptosis of ES cells in a dose-dependent manner and is expected to be a potential therapeutic agent for ES.

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