HeLa Research Articles

6615 Clinical features and genetic etiology of primary ovarian insufficiency in 46,XX children

Abstract Disclosure: F. Wang: None. X. Yin: None. X. Lan: None. P. Li: None. Purpose: The clinical features of primary ovarian insufficiency in 46,XX children were analyzed, the pathogenic genes of POI were screened, and the genetic etiology of primary ovarian insufficiency in children 46,XX was clarified. Method: The clinical data of 46,XX POI patients were collected, including uterine and ovarian ultrasound, serum follicle-stimulating hormone (FSH), estrogen, anti-mullerian hormone (AMH), chromosomal karyotype analysis, adrenal and thyroid function, and immune function. Whole exome sequencing (WES) and CNV were used to screen POI candidate genes. Novel mutations such as MCM8 gene were used to verify cell function. HE staining, immunohistochemistry, cell fluorescence, and Tunnel method were used to observe the changes in ovarian tissue structure. Construction overexpression plasmids of Mutant MCM8 gene in Hela cells. Result: 1. Clinical features of 46,XX POI: The age of onset of the children was 11.9∼16.1 years old. In addition to POI, the clinical phenotype showed height growth retardation. Among them, 4 cases were short stature, 3 cases were growth hormone deficiency, and 5 cases were backward in bone age. All of them showed elevated FSH (>25 IU/L), and 7 cases had low serum AMH (<0.5 ng/ml). 2.The Screening of POI candidate genes and pathogenicity analysis: 5 of the 10 children carried the mutations of the pathogenic genes with POI, that is, 2 cases (sisters of the same family) carried two novel heterozygous mutations of MCM8 gene, one infant carried two novel heterozygous mutations of CYP19A1 gene, and one case with delayed puberty carried one homozygous mutation of ZSW1M7 gene. 3.The MCM8 mutant were functionally validated: The sisters carried two novel mutations, namely p.C242R and p.S445*. The elder sister (the proband) showed POI, short stature and tumor susceptibility, while the younger sister showed a trend of gradual increase in FSH level and ovarian decline. The follicles of the proband disappeared and fibrous tissue proliferation, the MCM8 expression was lower, and the positive rate of cell apoptosis was higher than normal control. The downregulation of Bax and Capase3 expression in mutant MCM8 of Hela cell was delayed compared with wild type, indicating the persistence of apoptosis. Conclusion: 1. The clinical characteristics of 46,XX POI in children are ovarian hypoplasia, often combined with height growth retardation, biochemical manifestations of hypergonadotropic hypogonadism, decreased serum AMH, and karyotype 46,XX. 2.46,XX POI genetic etiology analysis found that five known POI-related pathogenic genes, namely CYP19A1, MCM8, and ZSW1M7, in which five novel mutations were identified.3.Mutant MCM8 promotes ovarian apoptosis by activating the Bcl-2/Bax/Caspase 3 pathway. Keywords: Primary ovarian insufficiency, short stature, apoptosis Presentation: 6/3/2024

Proteomic analysis of HeLa cells after stable transfection with the Chlamydia trachomatis CT143 gene

BackgroundThe CT143 protein of Chlamydia trachomatis (Ct) is a key immunodominant antigen and candidate type-III secretion substrate. Although CT143 expression has not been detected in the cytosol of infected cells, it is known to interfere with the physiological behavior of HeLa cells. This study aims to investigate how the CT143 protein affects the protein expression profile of HeLa cells, providing a basis for further research into Ct’s pathogenic mechanisms. MethodsWe constructed a stably transfected HeLa cell line, pCD513B-1-CT143-HeLa, and a control cell line, pCD513B-1-HeLa. Protein expression profiles of these cell lines were determined using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Differentially expressed proteins were identified, constructed into a database, and verified using parallel reaction monitoring (PRM). Bioinformatics software facilitated the preliminary analysis of the biological functions of these differential proteins. ResultsA total of 221 host proteins were differentially expressed, with 68 upregulated and 153 downregulated. These variations influence the regulation of peptidase activity and are crucial in biological processes such as cell secretion and protease activity. Significant changes were noted in protein processing, alcohol dehydrogenase activity, Aldo-Keto reductase activity, and peptidase regulator activity. Furthermore, alterations were observed in cellular components like the plasma membrane and cell periphery. Pathways involving the hematopoietic system, glycosaminoglycan degradation, retinol metabolism, and cytochrome P450-mediated exogenous drug metabolism were notably affected. Indirect interactions among differentially expressed proteins included three key nodal proteins: C3, IFIT3, and IFIT1. ConclusionThe successful construction of a host differential protein expression profile was achieved through stable transfection of HeLa cells with the CT143 gene. The differential proteins identified are implicated in regulating various biological processes such as intracellular signal transduction, cell secretion, protein processing, hydrolysis, and enzyme activity. These findings suggest that the CT143 protein may influence the host cell’s biological behavior by altering host protein expression, potentially hindering Ct growth and development.

β-Cyclodextrin-Modified Laser-Induced Graphene Electrode for Detection of N6-Methyladenosine in RNA.

Laser-induced graphene (LIG) possesses characteristics of easy handling, miniaturization, and unique electrical properties. We modified the surface of LIG by electropolymerizing β-cyclodextrin (β-CD), which was used to immobilize antibodies on the electrode surface for highly sensitive detection of targets. N6-methyladenosine (m6A) is the most prevalent reversible modification in mammalian messenger RNA and noncoding RNA, influencing the development of various cancers. Here, β-CD was electropolymerized to immobilize the anti-m6A antibody, which subsequently recognized the target m6A. This was integrated into the catalytic hydrogen peroxide-hydroquinone (H2O2-HQ) redox system using phos-tag-biotin to generate electrochemical signals from streptavidin-modified horseradish peroxidase (SA-HRP). Under optimal conditions, the biosensor exhibited a linear range from 0.1 to 100 nM with a minimum detection limit of 96 pM. The method was successfully applied to the recovery analysis of m6A from HeLa cells through spiking experiments and aims to inspire strategies for point-of-care testing (POCT).

A Carbazole-based Fluorescent Probe with AIE Performance and a Large Stokes Shift for Peroxynitrite Detection and Imaging in Live Cells.

A carbazole-based fluorescent probe YCN with AIE performance and a large Stokes shift (242nm) shift was synthesized by attaching 4-acetonitrile pyridine to the 3-phenylaldehyde butylcarbazole. Its structure was characterized by 1H NMR, 13C NMR and MS. Probe YCN has high selectivity and sensitivity toward ONOO-. The addition of ONOO- to the probe YCN solution results in a noticeable color change from pale yellow to colorless under natural light, and a fluorescent color change from bright orange-yellow to bright yellow-green under a 365nm UV lamp, which can be distinguished by the naked eye. The research results on the reaction mechanism showed that when YCN reacted with ONOO-, -C = C- was oxidized and broken into -CHO, and the ICT effect was significantly inhibited, resulting in changes in UV absorption and fluorescence emission phenomenon. The recognition mechanism was verified by 1H NMR, mass spectrometry (MS) and density function theory (DFT) calculations. The experiments of live cells imaging suggested that compound YCN can be used as a fluorescent probe for the detection of ONOO- in HeLa cells. This result indicates that YCN has potential application prospects in the biological aspects.

A Novel Lactam and Two Rare Mycophenolic Acid Derivatives from the Fungus Penicillium sclerotiorum JBHL321.

A novel lactam penicillactam (1), two rare mycophenolic acid (MPA) derivatives penimycophens A and B (2 and 3), together with two known biogenetically related MPA derivatives (4 and 5) and a known alkaloid (6) were isolated from the Penicillium sclerotiorum JBHL321. The structures of these compounds were determined by comprehensive spectroscopic analyses. The absolute configuration of 1 was confirmed by electronic circular dichroism (ECD) calculations. Compound 1 represent the rare example of a oxygen bridge-linked lactam from natural products. Structurally, compounds 2 and 3 were rare MPA derivatives featuring a methoxy group at C-3. The inhibitory activity of compounds 1-6 against two phytopathogenic fungi, three phytopathogenic bacteria and four cancer cell lines were evaluated. Compounds 2-5 exhibited significant cytotoxic activity against HeLa, MCF-7, A549 and MGC-803 cells with IC50 values in the low micromolar to nanomolar. Compound 3 exhibited especially cytotoxic activity against four different cell lines with IC50 values ranging from 0.06 to 0.14 μM, compared to IC50 values ranging from 0.62 to 2.51 μM for epirubicin.

Relative biological effectiveness of clinically relevant photon energies for the survival of human colorectal, cervical, and prostate cancer cell lines

Objective.Relative biological effectiveness (RBE) differs between radiation qualities. However, an RBE of 1.0 has been established for photons regardless of the wide range of photon energies used clinically, the lack of reproducibility in radiobiological studies, and outdated reference energies used in the experimental literature. Moreover, due to intrinsic radiosensitivity, different cancer types have different responses to radiation. This study aimed to characterize the RBE of clinically relevant high and low photon energiesin vitrofor three human cancer cell lines: HCT116 (colon), HeLa (cervix), and PC3 (prostate).Approach.Experiments were conducted following dosimetry protocols provided by the American Association of Physicists in Medicine. Cells were irradiated with 6 MV x-rays, an192Ir brachytherapy source, 225 kVp and 50 kVp x-rays. Cell survival post-irradiation was assessed using the clonogenic assay. Survival fractions were fitted using the linear quadratic model, and survival curves were generated for RBE calculations.Main results.Cell killing was more efficient with decreasing photon energy. Using 225 kVp x-rays as the reference, the HCT116 RBESF0.1for 6 MV x-rays,192Ir, and 50 kVp x-rays were 0.89 ± 0.03, 0.95 ± 0.03, and 1.24 ± 0.04; the HeLa RBESF0.1were 0.95 ± 0.04, 0.97 ± 0.05, and 1.09 ± 0.03, and the PC3 RBESF0.1were 0.84 ± 0.01, 0.84 ± 0.01, and 1.13 ± 0.02, respectively. HeLa and PC3 cells had varying radiosensitivity when irradiated with 225 and 50 kVp x-rays.Significance.This difference supports the notion that RBE may not be 1.0 for all photons through experimental investigations that employed precise dosimetry. It highlights that different cancer types may not have identical responses to the same irradiation quality. Additionally, the RBE of clinically relevant photons was updated to the reference energy of 225 kVp x-rays.

PAIP1 binds to pre-mRNA and regulates alternative splicing of cancer pathway genes including VEGFA

BackgroundPoly (A) binding protein interacting protein 1 (PAIP1) has been shown to causally contribute to the development and progression of cancer. However, the mechanisms of the PAIP1 regulation in tumor cells remain poorly understood.ResultsHere, we used a recently developed UV cross-linking and RNA immunoprecipitation method (iRIP-seq) to map the direct and indirect interaction sites between PAIP1 and RNA on a transcriptome-wide level in HeLa cells. We found that PAIP1 not only binds to 3’UTRs, but also to pre-mRNAs/mRNAs with a strong bias towards the coding region and intron. PAIP1 binding sites are enriched in splicing enhancer consensus GA-rich motifs. RNA-seq analysis revealed that PAIP1 selectively modulates the alternative splicing of genes in some cancer hallmarks including cell migration, the mTOR signaling pathway and the HIF-1 signaling pathway. PAIP1-regulated alternative splicing events were strongly associated with PAIP1 binding, demonstrating that the binding may promote selection of the nearby splice sites. Deletion of a PAIP1 binding site containing seven repeats of GA motif reduced the PAIP1-mediated suppression of the exon 6 inclusion in a VEGFA mRNA isoform. Proteomic analysis of the PAIP1-interacted proteins revealed the enrichment of the spliceosome components and splicing factors.ConclusionsThese findings suggest that PAIP1 is both a polyadenylation and alternative splicing regulator, that may play a large role in RNA processing via its role in alternative splicing regulation.

Effects of Zinc Phthalocyanine Photodynamic Therapy on Vital Structures and Processes in Hela Cells.

This work presents results on the efficiency of newly designed zinc phthalocyanine-mediated photodynamic therapy of both tumoral and nontumoral cell models using the MTT assay. Further detailed examinations of mechanistic and cell biological effects were focused on the HELA cervical cancer cell model. Here, ROS production, changes in the mitochondrial membrane potential, the determination of genotoxicity, and protein changes determined by capillary chromatography and tandem mass spectrometry with ESI were analyzed. The results showed that, in vitro, 5 Jcm-2 ZnPc PDT caused a significant increase in reactive oxygen species. Still, except for superoxide dismutase, the levels of proteins involved in cell response to oxidative stress did not increase significantly. Furthermore, this therapy damaged mitochondrial membranes, which was proven by a more than 70% voltage-dependent channel protein 1 level decrease and by a 65% mitochondrial membrane potential change 24 h post-therapy. DNA impairment was assessed by an increased level of DNA fragmentation, which might be related to the decreased level of DDB1 (decrease in levels of more than 20% 24 h post-therapy), a protein responsible for maintaining genomic integrity and triggering the DNA repair pathways. Considering these results and the low effective concentration (LC50 = 30 nM), the therapy used is a potentially very promising antitumoral treatment.

2-Ferrocene-4-amine-1H-benzo[d]imidazole and their derivatives: Synthesis, biological and docking study

An organometallic series based on 2-ferrocene-4-amine-1H-benzo[d]imidazole derivative has been synthesized and characterized through FTIR, H/CNMR, and LCMS. Their antimicrobial activity was assessed by testing them against B. subtills, Staphylococcus aureus, E. faecalis, S. marcescens, E. coli, K. pneumoniae, C. albicans, A. niger, Mucor sp. where they showed good zone of inhibition (ZOI) as compared to standards. The cytotoxicity of synthesized compounds was tested against Hela (cervical) and MCF-7 (breast) cancer cell lines where compound 5a and 5b displayed equivalent potency to the standard while 5e demonstrated highest cytotoxicity against both cell lines. Their comparative docking analysis with DNA suggested that docking score of 5e was −7.4 kcal/mol versus −6 kcal/mol of ethidium bromide, indicating that our designed compound acts via DNA intercalation.

A-297 Feasibility of a novel Seegene Multiplex PCR panel for the simultaneous detection of HSV-1, HSV-2, VZV and Mpox: a focus on inclusion, confirmation and LOD assessment

Abstract Background Characterized by atypical presentations of short prodromal symptoms, variations in clinical staging, non-uniform presentation of rash, and varied syndromes of proctitis and pharyngitis, the WHO declared the 2022 Mpox outbreak a global public health emergency in July 2022. The overlap of clinical presentation with other causes of vesicular lesions identified gaps in the diagnostic tests available for MPXV and highlighted the need for a multiplex PCR approach to the diagnosis of HSV, VZV, and MPXV among patients presenting with vesicular lesions. Currently, no FDA cleared multiplex approach offering a single unified approach to the diagnosis of HSV, VZV, and MPXV from a single sample exists. Multiplex assays offer a role in diagnostic support where clinicians are challenged to determine the causative agent because of overlap. Furthermore, no assay capable of providing MPXV clade II reporting, needed for compliance to select agent reporting rules, exist for routine clinical labs. This study outlines the initial feasibility of Seegene’s multiplex HSV, VZV, MPXV PCR assay, highlighting the LOD assessment from the 1st WHO International Standards, ZeptoMetrix® NATtrolTM Controls, and custom plasmids. Methods Gene targets representing conserved, differentiated regions of HSV-1, HSV-2, VZV, MPXV, and MPXV clade II were identified through phylogenetic analysis. A total of 28 oligonucleotide combinations, were applied to Seegene’s proprietary amplification and detection process creating a multiplex assay capable of 8 unique detection signals in 5 channels using the BioRad Opus® 96. 1st WHO International Standards (IS) for HSV-1/2, VZV, ZeptoMetrix® NATtrolTM and custom designed plasmids controls were titrated and spiked into Copan UTM supplemented with 5% HeLa cells and used to define target LOD panels ranging in concentrations tested with multiple replicates. Sample processing, DNA extraction, and PCR amplification were performed on the Seegene Starlet® automated liquid handler and BioRad® OPUS 96 using Seegene software. Results Detection rates for HSV-1 at target concentrations of 1x102 IU/ml (WHO) and 6.17x100 copies/ml (NATtrolTM) were 100%. Detection of HSV-1 with plasmid DNA at target concentrations of 1.25x101 was 95%. Detection rates for HSV-2 were 100% at 5x102 IU/ml (WHO) and 5.56x101 copies/ml (NATtrolTM). Detection of HSV-2 with plasmid DNA at target concentrations 2.5E1 was 100%. For VZV, detection rates of 100% were observed at LOD challenge pools comprising 1x102 IU/ml (WHO) and 1.85x101 copies/ml (NATtrolTM). Detection of VZV with plasmid DNA at target concentrations 2.5x101 was 95%. For MPXV, detection rates of 100% were observed when challenged with LOD pools at 1.67x101 copies/ml respectively. Detection of MPXV with plasmid DNA at target concentrations 6.25x100 was 95%. Confirmation of clade II reporting for MPXV was determined using phylogenetic analysis. Conclusions Standardize of Seegene’s multiplex HSV-1, HSV-2, VZV, MPXV assay using traceable quantitated standard resulted in >95% detection rates of 6.17x100 copies/ml for HSV-1, 5.56x101 copies/ml for HSV-2, 1.85x101 copies/ml for VZV, and 1.67x101 copies/ml for MPXV respectively. A single multiplex PCR assay offers advantages for the simultaneous detection of HSV, VZV, and MPXV DNA in lesions specimens where clinical features present overlap and in patients whose clinical history is difficult to obtain.

Design, synthesis and biological evaluation of thienopyridine derivatives as c-Met kinase inhibitors.

With cabozantinib as the precursor, a novel small molecule inhibitors of c-Met kinase with thieno [2,3-b] pyridine as the scaffold were designed, synthesized and evaluated for their biological activity against A549, Hela and MCF-7 cell lines. The in vitro activities of 16 compounds were tested by MTT method with cabozantinib as control drug. Most compounds had moderate to strong inhibitory activities on cells. Among them, compound 10 had the strongest inhibitory activity, which was superior to the lead compound cabozantinib. Its IC50 values for A549, Hela and MCF-7 cells were 0.005, 2.833 and 13.581μM, respectively. The colony formation assay demonstrated that compound 10 significantly inhibited the colony formation of A549 cells and suppressed their growth in a concentration-dependent manner. The wound healing assay showed that compound 10 could effectively inhibit the migration of cancer cells compared to a blank control group. The AO/EB assay demonstrated that compound 10 possesses the capability to effectively trigger apoptosis in a concentration-dependent manner. The elementary structure-activity relationship, molecular docking and pharmacokinetics studies revealed the significance of thieno [2,3-b] pyridine derivatives in anti-tumor activity.

Hydroxyapatite layer evolution on silica-based (45S10P) bioactive glass-ceramic nanoparticles doped with Zn2+ ions: Augmentation of in vitro bioactivity, antibacterial activity and textural modification for bone regeneration

Zinc-incorporated bioactive glasses (BG) and bioactive glass-ceramic (BGC) nanoparticles (BGC-NPs) represent highly adaptable, biodegradable, and bioactive materials in tissue engineering and regenerative medicine. This study delves into the synthesis, characterization and biomedical implications of Zn2+ ions-doped 45S10P spherical BGC-NPs, employing analytical techniques and in vitro assays. The resulting (Zn2++ 45S10P) BGC-NPs, prepared using a modified Strober's method, exhibited spherical morphology with excellent dispersion, as verified by TEM, FE-SEM, and DLS analyses. XRD, FTIR, and FESEM analysis revealed an augmented hydroxyapatite (HAp) layer formation with increasing zinc content. Zeta potential analysis showcased a shift from negative to positive values after immersion in simulated body fluid (SBF), indicating the bioactive potential of the developed BGC-NPs. Hemocompatibility assays indicated the biocompatibility of all BGC-NPs, demonstrating minimal hemolytic effects with incorporating Zn2+ ions below a standard threshold (<5 % lysis). Further, the migration assay revealed the potentiality of the BGC-NPs to stimulate the migration of HeLa cells. Cell viability assays employing MG-63 osteoblast-like cells highlighted enhanced cell viability over time, underscoring their non-toxic nature and potential for tissue regeneration. Antibacterial assays displayed significant inhibitory effects against Gram-negative compared to Gram-positive bacteria, underscoring their potential for anti-infection applications. Overall, the results affirm the promising prospects of Zinc incorporation into 45S10P BGC-NPs for bone tissue regeneration applications.

Molecular Actions of Enicostemma hyssopifolium Whole Plant Extract on HPV18-Infected Human Cervical Cancer (HeLa) Cells.

Enicostemma hyssopifolium (E. hyssopifolium) contains several bioactive compounds with anti-cancer activities. This study was performed to investigate the molecular effects of E. hyssopifolium on HPV18-containing HeLa cells. The methanol extract of E. hyssopifolium whole plant was tested for cytotoxicity by MTT assay. A lower and higher dose (80 and 160 μg/mL) to IC50 were analyzed for colonization inhibition (Clonogenic assay), cell cycle arrest (FACS analysis), and induction of apoptosis (AO/EtBr staining fluorescent microscopy and FACS analysis) and DNA fragmentation (comet assay). The HPV 18 E6 gene expression in treated cells was analyzed using RT-PCR and qPCR. A significant dose-dependent anti-proliferative activity (IC50 - 108.25±2 μg/mL) and inhibition of colony formation cell line were observed using both treatments. Treatment with 80 μg/mL of extract was found to result in a higher percent of cell cycle arrest at G0/G1 and G2M phases with more early apoptosis, while 160 μg/mL resulted in more cell cycle arrest at SUBG0 and S phases with late apoptosis for control. The comet assay also demonstrated a highly significant increase in DNA fragmentation after treatment with 160 μg/mL of extract (tail moments-19.536 ± 17.8), while 80 μg/mL of extract treatment showed non-significant tail moment (8.152 ± 13.0) compared to control (8.038 ± 12.0). The RT-PCR and qPCR results showed a significant reduction in the expression of the HPV18 E6 gene in HeLa cells treated with 160 μg/mL of extract, while 80 μg/mL did not show a significant reduction. The 160 μg/mL methanol extract of E. hyssopifolium demonstrated highly significant anti-cancer molecular effects in HeLa cells.

Bio-based fabrication of palladium nanoparticles by using Aegle marmelos leaf extract for biomedical applications: Cytotoxicity, cell imaging studies in HeLa cells and antioxidant activity

This paper presents the outcome of medicinal plant-derived synthesis of nanomaterial. The results in the present study provide a simple, novel, sustainable, and economically noble process for the synthesis of palladium nanoparticles using leaf extract Aegle marmelos (A. marmelos). Fourier transformed infrared spectroscopy (FTIR) was used to verify the surface layer of synthesized palladium (ALE@Pd) nanoparticles (NPs) enriched with several kinds of phytochemicals. Transmission electron microscopy (TEM) revealed that the synthesized NPs possess a ball shape and the average diameter was 4.31 ± 1.89 nm. The synthesized NPs possess stability confirmed by UV–visible (UV–Vis) spectrophotometry. The face-centered cubic (FCC) crystal structure of the fabricated nanoparticles was confirmed by powder X-ray diffraction (PXRD). Fabricated ALE@Pd NPs are subjected to further testing for their cytotoxicity in HeLa cell lines, as well as for a comparative study of antioxidant activity between the A. marmelos leaves and prepared ALE@Pd NPs. The anti-carcinogenic activity and cell imaging in HeLa cells were tested, and the minimal inhibition concentration (MIC or IC50) has been determined and found to be around 46 µg/mL for HeLa cell lines. Fabricated palladium nanoparticles derived from A. marmelos have the potential to be employed as an environmentally friendly and cost-effective anticancer agent. Furthermore, synthesized ALE@Pd NPs possess higher antioxidant activity than A. marmelos leaves extract (ALE) due to the increased phenolic content connected to the surface of produced palladium NPs as a capping agent. The minimal inhibition concentration of synthesized ALE@Pd NPs was found to be ∼5 µg/mL.

Biological active compounds from edible mushroom: Effect on hela cells

Background: Mushrooms are unique organisms rich in proteins, carbohydrates, vitamins, and minerals, which allows them to be used not only as a source of nutrition but also in cosmetics and medicine. The fungus P. ostreatus possesses two strong enzymatic complexes – lignin-decaying and cellulose-decaying. Objective: The research aimed to study the antitumor activity of treated mm-wave mycelial extracts of mushrooms on the HeLa cell line. Research on edible mushrooms against tumors is very actual and important for the field of functional food science and bioactive compounds field. Methods: The effect of the fungal culture extract on the cyto-photometric and morphometric indices of the HeLa cell line culture was studied according to the nature of proliferation and transcription of malignant tissues. Using high-resolution liquid chromatography, the amino acid composition of acid-soluble proteins in mycelial extracts of a mushroom culture treated with mm waves was studied. Results: Extracts of P. ostreatus mycelial culture significantly suppressed the growth of proliferative activity of human tumor cultures and completely suppressed the mitotic activity of continuous HeLa cell cultures to 72 hours. Our study revealed a significant increase in glutamic and aspartic acids in mycelial culture extracts treated by mm-waves. Conclusion: Based on HPLC analysis, we suggest polyfunctional peroxidase as the candidate for the role of biologically active compound in the composition of aqueous extracts of the mycelial culture. So, the polyfunctional peroxidase plays an active role in the therapeutic properties of the oyster mushroom. Keywords: anticancer activity, ant-inflammation activity, mm-waves, mushroom`s, mycelial extracts, HPLC of acid-soluble proteins, HeLa cell line.

Rational design, synthesis, and anticancer evaluation of pyridine and substituted aryl linked 1,3,4-oxadiazole derivatives

A new series of Pyridine and substituted aryl linked 1,3,4-oxadiazole derivatives (7a-j) were designed, synthesized and evaluated for their anticancer activities against HeLa, MDA-MB-231, MCF-7, and A549 cell lines by using standard MTT reduction assay protocol with doxorubicin as standard drug. It is very important to mention that all these synthesized derivatives do not show any cytotoxicity on HEK-293 kidney cell lines. Among them, the compound 7d showed better activity against HeLa and MCF-7 cell lines with IC50 values of 1.8 ± 0.3 µM and 1.3 ± 0.11 µM.

Abutilon indicum-mediated green synthesis of NiO and ZnO nanoparticles: Spectral profiling and anticancer potential against human cervical cancer for public health progression

BackgroundIntegrating nanomedicines for targeted cancer treatment and pursuing medicinally valuable components from nature are crucial for sustainable, potent alternatives to synthetic drugs in combating fatal diseases like cancer. Hence, a green synthesis of nickel oxide (NiO NPs) and zinc oxide nanoparticles (ZnO NPs) has been carried out by using the leaf extract of medicinally important plant Abutilonindicum. This sustainable approach to medical developments not only reduces the environmental effect of standard synthesis methods and offers new options for novel cancer therapeutics, but it also advances public health by using natural resources in a sustainable manner. MethodsThe synthesized nanoparticles were characterized by employing spectro-analytical techniques like UV–vis, FT-IR, SEM and powder XRD. Synthesized nanparticles were evaluated in the human cervical cancer cells (HeLa). ResultsNi-O stretching vibrations were observed at 402 cm−1, whereas that of Zn-O stretching was observed at 409 cm−1in the FT-IR spectrum, confirming the formation of nanoparticles. The XRD pattern revealed the crystallite size range of 1.35–2.84 nm for NiO NPs and 7.71–56.80 nm for ZnO NPs. The morphology of the nanoparticles, as indicated by the SEM images, was rod-like for NiO NPs and rock-shaped for ZnO NPs. Further, the cancer cell growth inhibition activity of the nanoparticles was examined by MTT assay against human cervical cancer cells (HeLa) proliferation and compared with cisplatin. MTT assay elucidated the significant anticancer efficacy of the synthesized nanoparticles, showcasing low IC50 values of 29±0.5 µg/ml for NiO NPs and 32±0.7 µg/ml for ZnO NPs. Furthermore, the anticancer activity of the NiO NPs was investigated using the Trypan blue dye exclusion technique, emphasizing the pronounced cytotoxic impact of NiO NPs on cancer cell viability. The outcomes underscore the notable anticancer properties of plant extract mediated metal nanoparticles as promising contenders for advancing cancer treatment modalities.

An Aggregation Induced Emission Binuclear Pt(II) Complex with Mitochondria Targeting and Cell Necrosis Inducing Capability

AbstractIn this study, we synthesized a novel binuclear Pt(II) derivative with aggregation‐induced emission (AIE) property (DPT) through a straightforward route. The IC50 of DPT in the Hela cell line was calculated to be 44.3±1.4 μM. Experimental data showed DPT could induce a decrease in mitochondrial membrane potential (MMP) and an increase in reactive oxygen species (ROS). Necrosis was observed in DPT‐treated Hela cells. In addition to the cytotoxicity, DPT also displayed mitochondrial imaging capability. Thus, this complex might potentially serve as a theragnostic reagent.

In vitro characterization of beta-cyclodextrin liposomal formulation of cisplatin nanomedicine

This study aims to develop a nanomedicine-based approach to protect healthy tissue during cancer treatment, improve patient compliance, reduce adverse drug reactions, and enhance the quality of life. By utilizing hydroxypropyl-β-cyclodextrin (BPCP) to increase drug hydrophilicity and liposomes for controlled release of cisplatin (CP), the hydroxypropyl-β-cyclodextrin-cisplatin liposomal formulation (BCDCPLP) demonstrated improved stability, targeted delivery, and reduced toxicity. The study confirmed that the above formulation had the highest cytotoxicity against HeLa cells, with significantly lower IC50 and IC90 values than cisplatin alone. This is attributed to improved drug stability and cellular uptake. Caspase-3 and cleaved PARP assays confirmed that BCDCPLP and CP-loaded liposomes induced substantial apoptosis, with BCDCPLP showing the highest increase in caspase-3 activity and PARP cleavage at 85 kDa. The ability of BCDCPLP to enhance apoptosis was further demonstrated through Western blot analysis, where the density of PARP cleavage bands was highest in BCDCPLP-treated cells, followed by CPLP and CP-treated cells. The study highlights that the cumulative release of PARP from CP-nanocarrier complexes leads to more efficient apoptosis than free CP. These findings align with the synergistic effects observed when combining CP with PARP inhibitors, which induce mitochondrial membrane permeabilization and cytochrome C release, activating the intrinsic apoptosis pathway. The potential of these nanomedicines, especially BCDCPLP, is in reducing the required dosage while maintaining therapeutic efficacy offers promising advances in cancer therapy.

Preparation and Evaluation of Polyethylene Glycol Conjugated Polyethylene Imine‐Coated Magnetic Nanoparticles for Paclitaxel Delivery to Cervical Cancer Cells

ABSTRACTThe purpose of this study was to synthesize and characterize the therapeutic effect of paclitaxel‐loaded polyethylene glycol modified polyethylene imine‐coated iron oxide nanoparticles on cervical cancer cell line (HeLa cell). The physiochemical characteristics of nanocarriers were confirmed by using several methods including X‐ray diffraction (XRD), Thermogravimetric analysis (TGA), Fourier transform infrared spectroscopy (FT‐IR), Vibrating sample magnetometer (VSM), Dynamic light scattering (DLS), and Transmission electron microscopy (TEM). The results showed the magnetic nanoparticles with size of 35 nm and excellent magnetic properties could load paclitaxel with capacity (16%) and encapsulation efficiency (80%) that has revealed pH‐dependent release behavior. The anticancer effects of nanoparticles were investigated using MTT assay. In comparison to free drug, paclitaxel‐loaded magnetic nanocarrier revealed more cytotoxicity against HeLa cell line and also resulted in an increased rate of apoptotic cancer cells, assessed by flow cytometry analysis. Furthermore, the developed magnetic nanocarrier showed suitable hemocompatibility which can be used in clinical applications.