Human Cervical Carcinoma Cells Research Articles

Crizotinib-induced anti-cancer activity in human cervical carcinoma cells via ROS-dependent mitochondrial depolarization and induction of apoptotic pathway.

Cervical cancer is one of the leading causes of mortality among women population worldwide. In spite of recurrent screening, vaccination, and chemotherapeutic interventions, combating cervical cancer still remains a challenge. Crizotinib is a small molecule inhibitor that targets mesenchymal epithelial transition factor (c-MET) and has been successfully studied for its anti-cancer effects in non-small cell lung cancer, pancreatic, gastric, renal, prostate, and breast carcinomas. Although c-MET is a well-known prognostic, diagnostic, and therapeutic target in cervical cancer, anti-cancer properties of its inhibitor crizotinib against cervical carcinoma, has not been explored yet. In the present study, the anti-cancer effects of crizotinib on cervical cancer cells were evaluated using various in vitro cell-based assays, such as labelling drug-treated cells with MTT, H2 DCFDA, Annexin V5-fluorescein isothiocyanate (FITC) antibody, JC-1, PI, and analysis using fluorescence-activated cell sorting (FACS). The molecule was found to effectively inhibit proliferation of cervical cancer cells HeLa and SiHa with an IC50 of 0.641 ± 0.0724 and 0.871 ± 0.104 μM, respectively, and induce apoptosis in a dose-dependent manner. Further investigations showed that crizotinib-induced production of reactive oxygen species (ROS) with increasing concentrations further resulted in mitochondrial membrane depolarization. However, the drug had no effect on cell cycle progression of HeLa and SiHa cells. Thus, the study elucidates the cytotoxic effects of crizotinib in cervical cancer cells by activation of ROS-dependent apoptotic pathway via mitochondrial depolarization. These findings will further aid the evaluation of other molecular mechanisms of crizotinib and would pave the way for its implication as a chemotherapeutic option in cervical cancer.

Itraconazole Increases Resolvin E3 Concentration and 12/15-lipoxygenase Inhibitor Attenuates Itraconazole Cytotoxicity in Cervical Cancer Cells

The anticancer mechanism of itraconazole remains unsolved; therefore, we studied itraconazole-induced alterations in specialized pro-resolving mediators (SPMs) in cancer cells. The human cervical squamous carcinoma cell line CaSki was cultured with or without 1 μM itraconazole. Liquid chromatography/mass spectrometry analysis was conducted to identify SPMs that were influenced by itraconazole. Cell growth experiments were conducted using itraconazole and inhibitors targeting the metabolic pathways of candidate SPMs. Resolvin E3, resolvin E2, prostaglandin J2 (PGJ2), delta-12-PGJ2, and maresin 2 were identified as candidate SPMs. The 12/15-lipoxygenase inhibitor, which is involved in the conversion of 18-hydroxy-eicosapentaenoic acid to resolvin E3, attenuated the inhibitory effect of itraconazole. Inhibition of the PGJ2 metabolic pathway did not interfere with itraconazole treatment. The metabolic pathway of SPMs, including resolving E3, could be proposed as an anticancer target of itraconazole.

Molecular Structural Evolution of Near-Infrared Cationic Aggregation-Induced Emission Luminogens: Preclinical Antimicrobial Pathogens Activities and Tissues Regeneration

Molecular Structural Evolution of Near-Infrared Cationic Aggregation-Induced Emission Luminogens: Preclinical Antimicrobial Pathogens Activities and Tissues Regeneration

Lipoic Acid-Modified Oligoethyleneimine-Mediated miR-34a Delivery to Achieve the Anti-Tumor Efficacy.

MiR-34a, an important tumor suppressor, has been demonstrated to possess great potential in tumor gene therapy. To achieve the upregulation of miR-34a expression level, an oligoethyleneimine (OEI) derivative was constructed and employed as the carrier through the modification with lipoic acid (LA), namely LA-OEI. In contrast to OEI, the derivative LA-OEI exhibited superior transfection efficiency measured by confocal laser scanning microscopy and flow cytometry, owing to rapid cargo release in the disulfide bond-based reduction sensitive pattern. The anti-proliferation and anti-migration effects were tested after the miR-34a transfection to evaluate the anti-tumor response, using human cervical carcinoma cell line HeLa as a model. The delivery of LA-OEI/miR-34a nanoparticles could achieve obvious anti-proliferative effect caused by the induction of cell apoptosis and cell cycle arrest at G1 phase. In addition, it could inhibit the migration of tumor cells via the downregulation of MMP-9 and Notch-1 level. Overall, the LA-OEI-mediated miR-34a delivery was potential to be used as an effective way in the tumor gene therapy.

Squamins C–F, four cyclopeptides from the seeds of Annona globiflora

Four cyclic octapeptides, squamins C–F, were isolated from the seeds of Annona globiflora Schltdl. These compounds share part of their amino acid sequence, -Pro-Met(O)-Tyr-Gly-Thr-, with previously reported squamins A and B. Their structures were determined using NMR spectroscopic techniques together with quantum mechanical calculations (QM-NMR), ESI-HRMS data and a modified version of Marfey's chromatographic method. All compounds showed cytotoxic activity against DU-145 (human prostate cancer) and HeLa (human cervical carcinoma) cell lines. Clearly, A. globiflora is an important source of bioactive molecules, which could promote the sustainable exploitation of this undervalued specie.

Synthesis, spectroscopic and biological activity evaluation of Ni(II), Cu(II) and Zn(II) complexes of schiff base derived from pyridoxal and 4-fluorobenzohydrazide

A novel Schiff base ligand, 4-fluoro-N-((3-hydroxy-5-(hydroxymethyl)-2-methylpyridin-4-yl)methylene)benzohydrazide (PLFBH) was synthesized by condensationof pyridoxal and 4-fluorobenzohydrazide. Its complexes with Ni(II), Cu(II), and Zn(II) metal ionswere prepared and characterized by spectroscopic IR, 1H-NMR, UV, LC-MS, ESR, and powder XRD studies and by elemental analysis and thermal analysis, molar conductance, and magnetic susceptibility measurements. The results indicate the geometry of the complexes to be hexa coordinate distorted octahedral. Based on the electronic absorption and fluorescence emission spectra and viscosity studies, an intercalative mode of binding of the complexes with CT-DNA was suggested, which was also supported by DNA docking studies. The docking studies of metal complexes with DNA were carried out using Autodock 4.2. The in vitro anticancer assay for the Cu(II)-PLFBH complex was performed to assess the ability of the complex to inhibit human cell proliferation on HeLa human cervical carcinoma cells, MCF-7 human breast carcinoma cells, and A549 human lung carcinoma cells. The Cu(II)-PLFBH complex exhibited moderate to good inhibitory effect on the cancer cell lines studied. The complexes showed good cleavageability toward plasmid pBR322 DNA. The metal complexes were found to show good antibacterial activity against gram positive bacteria, Staphylococcus aureus and Bacillus cereus and gram negative bacteria Escherichia coli and Pseudomonas aeruginosa cultures,while the ligand showed marginal activity. Supplemental data for this article is available online at https://doi.org/10.1080/15257770.2021.1961271 .

The Role of the Hematopoietic Cell-Specific Protein 1-Associated Protein X-1 in Human Papillomavirus 16 E2-Induced Human Cervical Squamous Carcinoma Cell Apoptosis via a Mitochondria-Dependent Pathway

Objectives: Human papillomavirus 16 (HPV 16) E2 is a transcriptional regulator that plays a key role in regulating a variety of biological responses. Hematopoietic cell-specific protein 1-related protein X-1 (HAX-1) is a mitochondrial membrane protein, and the HAX-1 gene is involved in the occurrence, growth, invasion, and metastasis of various human malignant tumors. The purpose of this study was to investigate the relationships among HPV 16 E2, the role of HAX-1 gene, and the underlying intracellular apoptotic mechanism of human cervical squamous carcinoma cells (C33a and SiHa). Methods: In this study, HAX-1 expression was examined using real-time polymerase chain reaction, Western blot, and immunohistochemical staining analysis. Apoptosis of cells was assessed by flow cytometry. The mitochondrial function was assessed by the mitochondrial copy number, reactive oxygen species (ROS) generation, the mitochondrial membrane potential (ΔΨm), and mitochondrial morphology. Results: Our study demonstrated that the expression of the HAX-1 gene was significantly increased in human cervical carcinoma tissues relative to noncancerous cervix tissues. HPV 16 E2 inhibited HAX-1 protein expression. Overexpression of HAX-1 increased the mitochondrial copy number, decreased the production of ROS, and maintained the integrity of the mitochondrial membrane and morphology. So, enhanced expression of the HAX-1 gene could abrogate the HPV 16 E2-induced cell apoptosis. Conclusion: Therefore, these data support a mechanism that HAX-1 plays a crucial role in HPV 16 E2-induced human cervical squamous carcinoma cell apoptosis in a mitochondrial-dependent manner.

Evaluation of bioactive properties of Pleurotus ostreatus mushroom protein hydrolysate of different degree of hydrolysis

Edible mushrooms are natural sources for numerous bioactive properties which can protect human body from oxidative damages, hypertension, abnormal cell growth etc., The present study disclosed that, bioactive properties of mushroom proteins are significantly enhanced by adopting enzymatic hydrolysis. This study established the mushroom protein hydrolysates (MPHs), produced from freshly cultivated edible mushroom, Pleurotus ostreatus, could be the supreme source of both bioactive and nutraceutical compounds. Here, both unhydrolysed mushroom extract (UME) and proteinase K treated MPHs of different degrees of hydrolysis DH% (55%, 60%, 65%, 70% and 80%) under optimized condition were evaluated for antioxidative, angiotensin rennin converting enzyme (ACE) inhibitory and antiproliferative activities at different concentrations (2.5, 5 and 10 mg/ml). Among these, 65% DH at 10 mg/ml exhibited profound antioxidative (DPPH, ABTS, hydroxyl radical scavenging, metal chelating, linoleic acid auto-oxidation, FRAP, and reducing power) and ACE inhibitory activities. Maximum amount of total phenolic and flavonoid content was also obtained in this case. MTT assay confirmed the remarkable antiproliferative properties of MPHs against human cervical carcinoma cell line (HeLa). Presence of a wide range of phenolic (i.e., gallic, trans-cinnamic, caffeic, ferulic, aspartic and vanillic acid) and flavonoid (quercetin) compounds in both UME and MPHs were identified through HPLC.

Raman Spectroscopic technique as an effective method for detecting adulteration in an Ayurvedic fungal medicine Phansomba (Phellinus merrillii)

Phansomba (Phellinus merrillii) is a traditional fungal medicine used in the western parts of Maharashtra for tooth ache, excessive salivation in children. It has also exhibited prominent in vitro anticancer activity against human cervical carcinoma cell lines. Over the past 20 years it was observed that availability of this fungal medicine became scanty and irregular mainly due to overexploitation and deforestation and more often a ‘look alike’adulter-ants have been commonly sold by the name phansomba. Therefore, the au-thenticity of this medicine is in question. Hence, the samples sold under the name phansomba were collected from various localities of Wersten Ghats and screened using Raman spectroscopy initially at range between 268–1773cm-1and later at a narrow range between 487–1436cm-1. The spectra of these samples were compared with the spectra of genuine phansomba (Ph. merrillii) sample. Later on, after transformation the spectral data into a bina-ry matrix neighbor joining and maximum likelihood tree were constructed. Similarly, the data was subjected to multivariate analysis using PCA and MDS. The results of these analysis indicated a clear separation of genuine phansomba samples and the adulterated samples. It was further revealed that the RAMAN bands at 487, 528, 786, 892, 915, and 1436 cm-1 are charac-teristic of Phellinus, particularly Ph. merrillii. The study, therefore, suggested that with the use of RAMAN spectra adulteration may easily be identified in phansomba samples by screening them at a range of 487–1436cm-1. We therefore, suggest that this range may serve as a signature range for Phel-linus and such specific spectral ranges may also be established for other herbal drugs and a good data base may be created.

The role of ESCRT-III and Annexin V in the repair of cell membrane permeabilization by the nanosecond pulsed electric field

Exposure of living cells to intense nanosecond pulsed electric field (nsPEF) increases membrane permeability to small solutes, presumably by the formation of nanometer-size membrane lesions. Mechanisms responsible for the restoration of membrane integrity over the course of minutes after nsPEF have not been identified. This study explored if ESCRT-III and Annexin V calcium-dependent repair mechanisms, which play critical role in resealing large membrane lesions, are also activated by electroporation and contribute to the membrane resealing. The extent of membrane damage and the time course of resealing were monitored by the time-lapse imaging of propidium (Pr) uptake in human cervical carcinoma (HeLa) cells exposed to trains of 300-ns PEF. The removal of the extracellular Ca2+ slowed down the resealing, although did not prevent it. Recruitment of CHMP4B protein, a component of ESCRT-III complex, to the electroporated plasma membrane was not observed, thus providing no evidence for possible contribution of the macro-vesicle shedding mechanism. In contrast, silencing the AnxA5 gene impaired resealing and reduced the viability of nsPEF-treated cells. We conclude that Annexin V but not ESCRT-III was involved in the repair of HeLa cells permeabilized by 300-ns stimuli, but it was not the only and perhaps not the main repair mechanism.

Antioxidant and Anticancer Efficacies of Anethum graveolens against Human Breast Carcinoma Cells through Oxidative Stress and Caspase Dependency.

Anethum graveolens, belonging to the family Apiaceae, has been extensively used for medicinal and therapeutic purposes since long. Plants encompass rich number of effective constituents with less toxicity. Thus, nowadays, the attempts are being made to search plant constituents that can prevent and reverse the chronic diseases, such as cancer. In this study, an in vitro antioxidant and anticancer efficacies of Anethum graveolens (AG-ME) were studied on human breast (MCF-7), lung (A-549), and cervical (HeLa) carcinoma cell lines. The antioxidant efficacies of AG-ME were evaluated by total antioxidant, DPPH radical scavenging, H2O2 scavenging, and ferrous reducing antioxidant assays. Further, the anticancer potential of AG-ME was also determined against different cancer cell lines. The AG-ME exhibited strong antioxidant activities as observed by antioxidant assays. AG-ME also showed a dose-dependent anticancer/cytotoxic potential against MCF-7, A-549, and HeLa cell lines. The AG-ME-induced reduction in GSH and increase in SOD activities indicates the role of oxidative stress in AG-ME-induced MCF-7 cell death. The results also exhibited that AG-ME triggered ROS production and significantly reduced MMP level. Moreover, a dose-dependent increase in caspase-3 and caspase-9 activities suggests that the AG-ME-induced MCF-7 cell death is caspase-dependent. Together, the present study provides reasoning and reassurance for the uses of A. graveleons for medical purposes as an antioxidant and anticancer agent. Additional investigations are required to examine biological and anticancer activities under an in vivo system to discover a possible beneficial use of AG-ME against diseases.

Novel chiral naphthalimide-cycloalkanediamine conjugates: Design, synthesis and antitumor activity

A novel series of enantiopure naphthalimide-cycloalkanediamine conjugates were designed, synthetized and evaluated for in vitro cytotoxicity against human colon adenocarcinoma (LoVo), human lung adenocarcinoma (A549), human cervical carcinoma (Hela) and human promyelocytic leukemia cell lines (HL-60). The cytotoxicity of the compounds was highly dependent on size and relative stereochemistry of the cycloalkyl ring as well as length of the spacer. By contrast, any kind of enantioselection was observed for each pair of enantiomers. Flow cytometric analysis indicated that compounds 22 and 23 could effectively induce G2/M arrest in the four previous cell lines despite a mild apoptotic effect.

Synthesis and biological evaluation of naphthoquinone phenacylimidazolium derivatives

In order to expand structural diversity and improve antitumor efficiency, forty new naphthoquinone phenacylimidazolium derivatives were designed, synthesized and evaluated. Good synthetic yields were obtained under mild conditions using easily available starting materials. Cytotoxicity of these compounds was evaluated in vitro against a panel of human tumor cell lines: human breast carcinoma cell lines (MCF-7), human cervical carcinoma cell lines (HeLa), and human lung carcinoma cell lines (A549). Among them, the optimal compound 7m showed splendid antiproliferative activity with low to 50 nM IC50 values against MCF-7 and excellent selectivity of 256-fold compared with the normal cell lines L929. Compound 7m induced apoptosis in a dose-dependent manner. Further mechanism experiments showed that compound 7m dramatically inhibited the expression of survivin and activated the pro-apoptotic protein caspase-3. Our results indicated that the structural modification on the 1,3-substituents of naphthoquinone imidazoliums without 2-substituent is also promising to obtain new antitumor compounds.

4-Hydroxyl-oxoisoaporphine, one small molecule as theranostic agent for simultaneous fluorescence imaging and photodynamic therapy as type II photosensitizer.

Oxoisoaporphine (OA) is a plant phototoxin isolated from Menispermaceae, however, its weak fluorescence and low water solubility impede it for theranostics. We developed here 4-hydroxyl-oxoisoaporphine (OHOA), which has good singlet oxygen-generating ability (0.06), strong fluorescence (0.72) and improved water solubility. OHOA displays excellent fluorescence for cell imaging and exhibits light-induced cytotoxicity against cancer cell. In vitro model of human cervical carcinoma (HeLa) cell proved that singlet oxygen generated by OHOA triggered photosensitized oxidation reactions and exert toxic effect on tumor cells. The MTT assay using HeLa cells verified the low cytotoxicity of OHOA in the dark and high phototoxicity. Confocal experiment indicates that OHOA mainly distributes in mitochondria and western blotting demonstrated that OHOA induces cell apoptosis via the mitochondrial pathway in the presence of light. Our molecule provides an alternative choice as a theranostic agent against cancer cells which usually are in conflict with each other for most traditional theranostic agents.

A Novel BODIPY Quaternary Ammonium Salt-Based Fluorescent Probe: Synthesis, Physical Properties, and Live-Cell Imaging.

The development of biological fluorescent probes is of great significance to the field of cancer bio-imaging. However, most current probes within the bulky hydrophobic group have limited application in aqueous medium and restricted imaging under physiological conditions. Herein, we proposed two efficient molecules to study their physical properties and imaging work, and the absorption and fluorescence intensity were collected with varying ions attending in aqueous medium. We enhance the water solubility through the quaternization reaction and form a balance between hydrophilic and hydrophobicity with dipyrrome-theneboron difluoride (BODIPY) fluorophore. We introduced pyridine and dimethylaminopyridine (DMAP) by quaternization and connected the BODIPY fluorophore by ethylenediamine. The final synthesized probes have achieved ideal affinity with HeLa cells (human cervical carcinoma cell line) in live-cell imaging which could be observed by Confocal Microscope. The probes also have a good affinity with subcutaneous tumor cells in mice in in vivo imaging, which may make them candidates as oncology imaging probes.

The Newly Synthetized Chalcone L1 Is Involved in the Cell Growth Inhibition, Induction of Apoptosis and Suppression of Epithelial-to-Mesenchymal Transition of HeLa Cells.

Over the past decades, natural products have emerged as promising agents with multiple biological activities. Many studies suggest the antioxidant, antiangiogenic, antiproliferative and anticancer effects of chalcones and their derivatives. Based on these findings, we decided to evaluate the effects of the newly synthetized chalcone L1 in a human cervical carcinoma cell (HeLa) model. Presented results were obtained by western blot and flow cytometric analyses, live cell imaging and antimigratory potential of L1 in HeLa cells was demonstrated by scratch assay. In the present study, we proved the role of L1 as an effective agent with antiproliferative activity supported by G2/M cell cycle arrest and apoptosis. Moreover, we proved that L1 is involved in modulating Transforming Growth Factor-β1 (TGF-β) signal transduction through Smad proteins and it also modulates other signalling pathways including Akt, JNK, p38 MAPK, and Erk1/2. The involvement of L1 in epithelial-to-mesenchymal transition was demonstrated by the regulation of N-cadherin, E-cadherin, and MMP-9 levels. Here, we also evaluated the effect of conditioned medium from BJ-5ta human foreskin fibroblasts in HeLa cell cultures with subsequent L1 treatment. Taken together, these data suggest the potential role of newly synthesized chalcone L1 as an anticancer-tumour microenvironment modulating agent.

DNA BINDING, IN VITRO CYTOTOXICITY AND ANTICANCER DRUG MECHANISM OF COPPER(II) COMPLEX CONTAINING PYRIDYL-TRIAZINE LIGAND

Mononuclear copper(II) complex [Cu(dppt)2(H2O)2](ClO4)2 (1), where dppt is bidentate NpyNtz donor asymmetric ligand (pyridyl-triazine) has been isolated. The X-ray crystal structure of 1 possesses a CuN4O2 chromophore with elongated octahedral geometry. The electronic and EPR spectral properties demonstrate that the solvent molecules strongly interacted in the axial position thereby weakens the CuN4 coordination plane. Absorption and emission spectral and electrochemical measurements clearly show the partial intercalative binding of 1 to calf thymus (CT) DNA. Remarkably, it exhibits potent cytotoxicity (IC50, 3.73 M) against human cervical carcinoma cells (HeLa), which is 4.5 times better than cisplatin and is non-toxic (IC50, >500 M) to normal mouse embryonic fibroblasts cells (NIH 3T3). It blocks cell cycle progression of HeLa cells in G1 phase. FACSverse analysis of 1 is suggestive of ROS (reactive oxygen species) generation and absolutely induces apoptotic cell death in HeLa cells.

Luminescent, protein-binding and imaging properties of hyper-stable water-soluble silicon quantum dots

Silicon quantum dots (SiQDs) bear the advantages of photostability and biocompatibility, but their synthesis and purification are relatively complex, and explorations on their protein-binding behaviors are very limited. Herein, water-soluble amino-terminated SiQDs were firstly synthesized in the absence of any extra reductants or catalysts. The absorption of synthesized SiQDs covered the entire UV band with the maximum absorption wavelength around 290 nm. Both their powder and solution emitted strong blue fluorescence when excited by UV light of 365 nm. The luminescent property of SiQDS aqueous solution was stable under broad regions of acidity (pH 1–12) and temperature (0–80 °C), which was also not affected by proteins. The SiQDs could bind with human serum albumin (HSA) through hydrogen bonds, which made fluorescence resonance energy transfer and quenching occur. The interaction mechanisms were analyzed through fluorescence lifetime decay curves, UV spectra, thermodynamics, and molecular surface modeling. The structural alterations were systematically deciphered by 3D fluorescence and circular dichroism spectra. Meanwhile, the cytotoxicity and cell fluorescence imaging performance were investigated using human cervical carcinoma cell lines (HeLa). The results suggested an exciting prospect of as-synthesized SiQDs in fluorescence imaging and related biomedical applications.

Characterization of an Antiviral Component in Human Seminal Plasma.

Numerous types of viruses have been found in human semen, which raises concerns about the sexual transmission of these viruses. The overall effect of semen on viral infection and transmission have yet to be fully investigated. In the present study, we aimed at the effect of seminal plasma (SP) on viral infection by focusing on the mumps viral (MuV) infection of HeLa cells. MuV efficiently infected HeLa cells in vitro. MuV infection was strongly inhibited by the pre-treatment of viruses with SP. SP inhibited MuV infection through the impairment of the virus’s attachment to cells. The antiviral activity of SP was resistant to the treatment of SP with boiling water, Proteinase K, RNase A, and DNase I, suggesting that the antiviral factor would not be proteins and nucleic acids. PNGase or PLA2 treatments did not abrogate the antiviral effect of SP against MuV. Further, we showed that the prostatic fluid (PF) showed similar inhibition as SP, whereas the epididymal fluid and seminal vesicle extract did not inhibit MuV infection. Both SP and PF also inhibited MuV infection of other cell types, including another human cervical carcinoma cell line C33a, mouse primary epididymal epithelial cells, and Sertoli cell line 15P1. Moreover, this inhibitory effect was not specific to MuV, as the herpes simplex virus 1, dengue virus 2, and adenovirus 5 infections were also inhibited by SP and PF. Our findings suggest that SP contains a prostate-derived pan-antiviral factor that may limit the sexual transmission of various viruses.

ISOLATION OF PROTEIN FROM THE SPINE VENOM OF PTEROIS VOLITANS FOUND IN THE INDONESIAN OCEAN, USING A HEATING PROCESS, FOR ANTICANCER, ANTIRETROVIRAL, ANTIBACTERIAL, AND ANTIOXIDANT ASSAYS

Objective: This research investigates the antibacterial, anticancer, antioxidant, and antiretroviral activities of the lionfish spine poison extract.
 Methods: Isolation and purification of the phospholipase A2 (PLA2) protein obtained from the spine poison were conducted through the following stages, including, extraction of the venom by sonication, heating, and purification using gradual saturation levels of ammonium sulfate. Furthermore, the purity and concentration of PLA2 were analyzed using the Lowry test and Marinetti’s method, respectively, while its protein content was ascertained through SDS-PAGE. Toxicity was then evaluated employing the brine shrimp lethality test (BSLT), and its anticancer activity was assessed in human cervical carcinoma cells (HeLa cells). Finally, its antioxidant, antibacterial, and antiretroviral activities were analyzed using the DPPH method, agar diffusion test against Salmonella sp. and E. coli, and SRV-2 and RT-qPCR tests, respectively.
 Results: The protein demonstrated 37.79% inhibition for anticancer activity, IC50 1312 ppm for antioxidant activity, 98.81%, and 89.28% inhibition of E. coli and Salmonella sp. respectively for antibacterial activity and 98.13% inhibition for antiretroviral activity.
 Conclusion: It can be concluded that lionfish (Pterois volitans) has the potential to be developed as an antioxidant, anticancer, antibacterial, and antiretroviral agent. Furthermore, the pharmacological activity of its spine venom was determined by isolating PLA2 protein from its extract, using an optimum heating temperature of 70 °C and an ammonium sulfate saturation level of 80%.