Human Cervical Cancer Cells Research Articles

Predictive Value and Immunological Role of the HSPA5 Gene in Cervical Cancer.

Cervical cancer (CC) ranks fourth among women's malignancies worldwide and seriously affects women's health. HSPA5 is a heat shock protein, also known as glucose regulatory protein 78 (GRP78). Upregulation of HSPA5 has been reported to be closely associated with multiple types of tumors. However, the specific role of HSPA5 in cervical cancer has not been discovered. In our study, we explored the prognostic value of HSPA5 in CC. Here, we analyzed the (TCGA) and (UCSC) databases, the analysis of HSPA5 in many tumors types was conducted with the "wilcox. test" method. A False Discovery Rate (FDR) value < 0.05 and Log2 | (fold change, FC) |> 1 were set as the cutoffs. "*", "**", and "***" indicate FDR < 0.05, < 0.01, and < 0.001, respectively, and further used human cervical cancer cells for q-PCR and western blotting detection. q-PCR and western blotting results showed that HSPA5 was highly expressed in cervical cancer cells, while it was expressed at low levels in normal cells (P < 0.05).We also analyzed the immunohistochemical data. immunohistochemical analysis results showed that HSPA5 was highly expressed in human cervical cancer, while it was expressed at low levels in normal tissues (P < 0.05). Analysis in TCGA-UCSC showed that the proportion of G3 in the group with high expression of HSPA5 was relatively high (P < 0.05). Enrichment analysis and survival analysis showed that the increased expression of HSPA5 in cervical cancer was related to the survival of CC and was involved in the regulation of biological behavior and molecular signaling pathways of cervical cancer. The correlation analysis of immune checkpoint and immune infiltration showed that HSPA5 was involved in the regulation of immune process of cervical cancer (P < 0.05). Drug sensitivity correlation analysis showed that HSPA5 was a sensitive target for tumor drugs (P < 0.05). In brief, those results suggest that HSPA5 can act as an oncogene of CC development and can serve as an effective predictive biomarker in cervical cancer.

Inclusion Complexes of Litsea cubeba (Lour.) Pers Essential Oil into β-Cyclodextrin: Preparation, Physicochemical Characterization, Cytotoxicity and Antifungal Activity.

The uses of natural compounds, such as essential oils (EOs), are limited due to their instability to light, oxygen and temperature, factors that affect their application. Therefore, improving stability becomes necessary. The objective of this study was to prepare inclusion complexes of Litsea cubeba essential oil (LCEO) with β-cyclodextrin (β-CD) using physical mixing (PM), kneading (KN) and co-precipitation (CP) methods and to evaluate the efficiency of the complexes and their physicochemical properties using ATR-FTIR, FT-Raman, DSC and TG. The study also assessed cytotoxicity against human colorectal and cervical cancer cells and antifungal activity against Aspergillus flavus and Fusarium verticillioides. The complexation efficiency results presented significant evidence of LCEO:β-CD inclusion complex formation, with KN (83%) and CP (73%) being the best methods used in this study. All tested LCEO:β-CD inclusion complexes exhibited toxicity to HT-29 cells. Although the cytotoxic effect was less pronounced in HeLa tumor cells, LCEO-KN was more active against Hela than non-tumor cells. LCEO-KN and LCEO-CP inclusion complexes were efficient against both toxigenic fungi, A. flavus and F. verticillioides. Therefore, the molecular inclusion of LCEO into β-CD was successful, as well as the preliminary biological results, evidencing that the β-CD inclusion process may be a viable alternative to facilitate and increase future applications of this EO as therapeutic medication, food additive and natural antifungal agent.

In Vitro Antiviral and Anticancer Effects of Tanacetum sinaicum Essential Oil on Human Cervical and Breast Cancer.

Cervical cancer has been linked to human papillomavirus (HPV) types 16 and 18. Essential oils (EOs) are vital natural products of plants with various therapeutic and biological properties. The purpose of this study is to investigate and assess Tanacetum sinaicum essential oil's possible antiviral and anticancer properties, with a focus on its in vitro effects on human cervical cancer and human breast adenocarcinoma cell lines. Tanacetum sinaicum EO was extracted via hydrodistillation (HD) and characterized using gas chromatography-mass spectrometry (GC-MS). MTT assay was used to determine the cell viability of Hela (a human epithelial cervical cancer) and MCF-7 (human breast adenocarcinoma) cell lines. Quantitative real-time polymerase chain reaction (PCR) was utilized to assess the antiviral efficacy of EO against HPV-16 and 18, and anti-metastatic characteristics. The biological activity of EO was assessed using Autophage and Cell genotoxicity via the comet assay. EO is mostly composed of chrysanthenyl acetate, thujone, and verbenol. The cell viability was reduced after 24 hours of incubation at doses from 100 to 400 µg/ml. Concentrations of 800 to 3,200 µg/ml significantly inhibit cell growth. After a 24-hour incubation period, doses ranging from 100 to 400 µg/ml reduced cell viability from 62 to 72%. Concentrations of 800 to 3,200 µg/ml significantly suppress cell growth by over 95%. In MCF7 and HeLa cell lines, EO lowered virus copy numbers in a dose-dependent manner, with higher concentrations of the oil inhibiting virus replication more effectively. EO treatment increased the number of autophagosomes/autolysosomes and acidic vesicular organelles in both cell lines. On the HeLa and MCF7 cell lines, EO demonstrated antiproliferative and antimetastatic effects. The results demonstrated that EO had dose-dependent genotoxic effects on both cancer cell lines, as evidenced by DNA damage. Tanacetum sinaicum EO is a prospective source of natural bioactive compounds that can be employed in pharmaceutical and medicinal applications due to its antiviral, antiproliferative, anti-metastatic and genotoxic properties.

FNDC5 inhibits malignant growth of human cervical cancer cells via restraining PI3K/AKT pathway.

Cervical cancer (CxCa) is the fourth most frequent cancer in women. This study aimed to determine the role and underlying mechanism of fibronectin type III domain-containing protein 5 (FNDC5) in inhibiting CxCa growth. Experiments were performed in human CxCa tissues, human CxCa cell lines (HeLa and SiHa), and xenograft mouse model established by subcutaneous injection of SiHa cells in nude mice. Bioinformatics analysis showed that CxCa patients with high FNDC5 levels have a longer overall survival period. FNDC5 expression was increased in human CxCa tissues, HeLa and SiHa cells. FNDC5 overexpression or FNDC5 protein not only inhibited proliferation, but also restrained invasion and migration of HeLa and SiHa cells. The effects of FNDC5 were prevented by inhibiting integrin with cilengitide, activating PI3K with recilisib or activating Akt with SC79. FNDC5 inhibited the phosphorylation of PI3K and Akt, which was attenuated by recilisib. PI3K inhibitor LY294002 showed similar effects to FNDC5 in HeLa and SiHa cells. Intravenous injection of FNDC5 (20 μg/day) for 14 days inhibited the tumor growth, and reduced the proliferation marker Ki67 expression and the Akt phosphorylation in the CxCa xenograft mouse model. These results indicate that FNDC5 inhibits the malignant phenotype of CxCa cells through restraining PI3K/Akt signaling. Upregulation of FNDC5 may play a beneficial role in retarding the tumor growth of CxCa.

Design, Synthesis, and In vitro Anti-cervical Cancer Activity of a Novel MDM2-p53 Inhibitor Based on a Chalcone Scaffold.

Several novel fluorinated chalcone derivatives were synthesized, and their in vitro anticervical cancer activity and mechanism of action were investigated using the parent nucleus of licorice chalcone as the lead compound backbone and MDM2-p53 as the target. In this study, 16 novel chalcone derivatives (3a-3r) were designed and synthesized by molecular docking technology based on the licorice chalcone parent nucleus as the lead compound scaffold and the cancer apoptosis regulatory target MDM2-p53. The structures of these compounds were confirmed by 1H-NMR, 13C-NMR, and HR-ESI-MS. The inhibitory effects of the compounds on the proliferation of three human cervical cancer cell lines (SiHa, HeLa, and C-33A) and two normal cell lines (H8 and HaCaT) were determined by MTT assay, and the initialstructure-activity relationship was analyzed. Transwell and flow cytometry were used to evaluate the effects of target compounds on the inhibition of cancer cell migration and invasion, apoptosis induction, and cell cycle arrest. Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot (WB) were used to detect the effects of candidate compounds on mRNA, p53, and Murine double minute 2 (MDM2) protein expression. The binding characteristics of the target compounds to the MDM2 protein target in the p53-MDM2 pathway were evaluated by molecular docking technology. The target compounds had considerable inhibitory activity on the proliferation of three cervical cancer cell lines. Among them, compound 3k (E)-3-(4-(dimethylamino)phenyl)-2-methyl-1-(3-(trifluoromethyl)phenyl) prop-2-en-1-one) showed the highest activity against HeLa cells (IC50=1.08 μmol/L), which was better than that of the lead compound Licochalcone B, and 3k showed lower toxicity to both normal cells. Compound 3k strongly inhibited the migration and invasion of HeLa cells and induced apoptosis and cell cycle arrest at the G0/G1 phase. Furthermore, compound 3k upregulated the expression of p53 and BAX and downregulated the expression of MDM2, MDMX, and BCL2. Moreover, molecular docking results showed that compound 3k could effectively bind to the MDM2 protein (binding energy: -9.0 kcal/mol). These results suggest that the compounds may activate the p53 signaling pathway by inhibiting MDM2 protein, which prevents cancer cell proliferation, migration, and invasion and induces apoptosis and cell cycle arrest in cancer cells. This study provides a new effective and low-toxicity drug candidate from licochalcone derivatives for treating cervical cancer.

Redox-Responsive Gold Nanoparticles Coated with Hyaluronic Acid and Folic Acid for Application in Targeting Anticancer Therapy.

Methotrexate (MTX) has poor water solubility and low bioavailability, and cancer cells can become resistant to it, which limits its safe delivery to tumor sites and reduces its clinical efficacy. Herein, we developed novel redox-responsive hybrid nanoparticles (NPs) from hyaluronic acid (HA) and 3-mercaptopropionic acid (MPA)-coated gold NPs (gold@MPA NPs), which were further conjugated with folic acid (FA). The design of FA-HA-ss-gold NPs aimed at enhancing cellular uptake specifically in cancer cells using an active FA/HA dual targeting strategy for enhanced tumor eradication. MTX was successfully encapsulated into FA-HA-ss-gold NPs, with drug encapsulation efficiency (EE) as high as >98.7%. The physicochemical properties of the NPs were investigated in terms of size, surface charges, wavelength reflectance, and chemical bonds. MTX was released in a sustained manner in glutathione (GSH). The cellular uptake experiments showed effective uptake of FA-HA-ss-gold over HA-ss-gold NPs in the deep tumor. Moreover, the release studies provided strong evidence that FA-HA-ss-gold NPs serve as GSH-responsive carriers. In vitro, anti-tumor activity tests showed that FA-HA-ss-gold/MTX NPs exhibited significantly higher cytotoxic activity against both human cervical cancer (HeLa) cells and breast cancer (BT-20) cells compared to gold only and HA-ss-gold/MTX NPs while being safe for human embryonic kidney (HEK-293) cells. Therefore, this present study suggests that FA-HA-ss-gold NPs are promising active targeting hybrid nanocarriers that are stable, controllable, biocompatible, biodegradable, and with enhanced cancer cell targetability for the safe delivery of hydrophobic anticancer drugs.

Alterations of UHRF family Expression and was regulated by High Risk Type HPV16 in Uterine Cervical Cancer

The altered protein expression of inverted CCAAT box-binding protein of 90 kDa/ubiquitin-like with PHD and RING finger domains 1 (ICBP90/UHRF1), and Np95-like ring finger protein (NIRF)/UHRF2, which belong to the ubiquitin-like with PHD and RING finger domains (UHRF) family, is linked to tumor malignancy and the progression of various cancers. In this study, we analyzed the UHRF family expression in cervical cancers, and it's regulation by human papillomavirus (HPV). Western blotting was performed to analyze protein expression in cervical cancer cell lines. Immunohistochemical analysis were used to investigate the expression of UHRF family and MIB-1 in cervical cancer tissues. Transfection were done for analyze the relationship between UHRF family and HPVs. We showed that NIRF expression was decreased and ICBP90 expression was increased in cervical cancers compared to normal counterparts. Western blotting also showed that NIRF expression was quite low levels, but ICBP90 was high in human cervical cancer cell lines. Interestingly, ICBP90 was up regulated by high risk type HPV16 E6 and E7, but not low-risk type HPV11. On the other hand, NIRF was down regulated by high risk type HPV16 E6 but not by E7. Low risk type HPV11 E6 did not affect the NIRF expression at all. We propose that ICBP90 overexpression, and reduced NIRF expression, found in cervical cancers, is an important event of a cervical carcinogenesis, and especially ICBP90 may offer a proliferating marker and therapeutic target for treating uterine cervical cancers.

Abstract 7290: Novel plasmid-based screening method for cervical cancer

Abstract Introduction: Alabama (AL) is tied for third-highest incidence rate and has second-highest death rate of cervical cancer as of 2020. Globally, cervical cancer has the fourth-highest incidence among cancers in women; 90% of deaths from cervical cancer in 2020 occurred in low- and middle-income (LMIC) countries. Inhibitor of differentiation/DNA binding 1 (Id1) expression correlates with tumorigenesis, clinical grade/stage, and aggressiveness of cervical cancer. Thus, Id1 expression could be exploited in novel screening approaches that aim to improve survival rates of cervical cancer in LMIC and clinically underserved settings. As a first step in developing an Id1-based screening approach, these preclinical studies utilized a plasmid that induces expression of a secreted reporter enzyme (secreted embryonic alkaline phosphatase, SEAP), controlled by the Id1 promoter sequence (pId1-SEAP). These studies examined relationships between Id1 expression and kinetics of reporter expression following plasmid transfection to detect human cervical cancer cells. Methods and Results: To confirm the relevance of Id1 as a target in cervical cancer, immunohistochemical staining for Id1 was performed on a human cervical cancer tissue microarray. Western blotting was performed to determine Id1 expression in human cervical cancer cell lines (HeLa, SiHa, CaSki). Optimal transfection conditions with pId1-SEAP were determined experimentally for each line, and then media was sampled through a 48-hour period post-transfection to quantify reporter SEAP expression. Immunohistochemical stain for Id1 showed that early stage and late stage human cervical cancer tissues expressed significantly (p &amp;lt; 0.05) higher levels of Id1 compared to normal tissues, with stain optical density values being 3e5±1e5 for early and late stage compared to 3e4±3e4 for normal tissues (early stage p=0.0001, late stage p=0.0002). Western blot for Id1 in HeLa, SiHa, and CaSki lines showed that all three positively expressed Id1, with Beta-actin-normalized expression values of 0.78±0.12, 1.12±0.05, and 0.76±0.08, respectively. The cell lines transfected with pId1-SEAP expressed SEAP at rates significantly higher than control cells, with the highest rates of production seen during the 36-48-hour post-transfection period. Rates of SEAP production (counts/hour) during this time period were 1e4±3e3 (p=0.0278), 8e2±1e2 (p=0.0834), and 3e2±5e1 (p=0.0683) for transfected HeLa, SiHa, and CaSki cells, respectively. Conclusions: The pId1-SEAP system enabled sensitive detection of cervical cancer cells. These preliminary data support future studies evaluating the in vivo performance of the system in preclinical models of early stage cervical cancer. Further development of this approach could enable sensitive and reliable detection of cervical cancer during early stages when treatment is most effective. Citation Format: Abbigael Eli, Yolanda Hartman, Benjamin B. Kasten, Rebecca Arend, Jason Warram. Novel plasmid-based screening method for cervical cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7290.

Abstract 2873: Radiation therapy promotes polyunsaturated fatty acid mediated ferroptosis in cervical cancer: Role of AMPK/xCT/GPX4 axis

Abstract Purpose: Previously, we found that mono-and di-unsaturated fatty acids can sensitize cervical cancer to radiotherapy (RT) through a p53 dependent apoptotic mechanism that is distinct from ferroptosis (Muhammad et al., Cancer Res. 2022; 82: 4515-27). Polyunsaturated fatty acids (PUFAs) enhance therapeutic response in preclinical tumor models by increasing lipid peroxidation (L-ROS) and induction of ferroptosis. Ferroptosis is a form of cell death that requires abundant cytosolic labile iron to promote membrane lipid peroxidation. The objective of the present study is to test whether the combination of RT + PUFAs promotes cell death via increased membrane lipid peroxidation and ferroptosis in cervical cancer, and to determine the impact of RT + PUFAs on cervix tumor cell metabolism and mitochondrial function. Experimental Procedures: Cell viability and clonogenic cell survival assays were performed using 8 human cervical cancer cell lines including normal cervix epithelial controls incubated with PUFAs (EPA, DHA and AA), before and after RT. Western blotting, siRNA-mediated knockdown, pharmacologic inhibition and activation were performed to determine whether ferroptosis dependent signals were mediating PUFA-induced RT sensitivity. Oxidative stress parameters were quantified using mass spectrometry and fluorometric dyes. Flow cytometry was used to quantify total cell ROS, lipid ROS, mitochondrial ROS and labile iron pool. Mitochondrial morphology and function were characterized using microscopy and the Seahorse assay. Summary: PUFAs + RT increased total ROS and LROS accumulation followed by a ferroptotic tumor cell death, which was iron-dependent and could be prevented by the addition of iron chelator and lipophilic antioxidants, Ferrostatin-1 and Liproxstatin-1. GPX4, a thiol dependent enzyme that eliminates lipid peroxides, and the cystine-glutamate antiporter (xCT) a key source of cystine for reduced glutathione (GSH) synthesis, were downregulated in response to PUFAs + RT. PUFAs + RT induced increased mitochondrial ROS and subsequent metabolic dysfunction. Mechanistically, the expression of AMPK, a key regulator of metabolic/energy homeostasis and mitochondrial biogenesis, was upregulated by PUFAs + RT. Pharmacological activation and inhibition of AMPK further enhanced and abrogated the effect of PUFA + RT, respectively. Conclusions: PUFAs modify the RT sensitivity of cervical cancer cells by inducing cell death through ferroptosis and mitochondrial dysfunction via AMPK activation. In vivo experiments are ongoing in immunocompromised and immunodeficient mouse tumor models to confirm this finding and determine the impact of PUFAs on RT induced tumor cell death and anti-tumor immunity. Citation Format: Naoshad Muhammad, Nishanth Gabriel, Louise M. Bengtsson, Tyler McKinnish, Naomi Msengi, Kevin Cho, Gary Patti, Julie K. Schwarz. Radiation therapy promotes polyunsaturated fatty acid mediated ferroptosis in cervical cancer: Role of AMPK/xCT/GPX4 axis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2873.

Abstract 1830: Bergenin inhibits growth of human cervical cancer cells by decreasing galectin-3 and MMP-9 expression

Abstract Cervical cancer is still the leading of cancer mortality worldwide, even after introducing a vaccine against Human papillomavirus (HPV), due to low vaccine coverage, especially in the developing world. Cervical cancer is primarily treated by Chemo/Radiotherapy, depending on the disease stage, with Carboplatin/Cisplatin-based drug regime. These drugs being non-specific, target rapidly dividing cells, including normal cells, so safer options are needed for more targeted therapy and lower off-target toxicity. Natural products offer an attractive option compared to synthetic drugs due to their well-established safety profile and capacity to target multiple oncogenic hallmarks of cancer like inflammation, angiogenesis, etc. In the current study, we investigated the effect of Bergenin (C-glycoside of 4-O-methylgallic acid), a natural polyphenol compound that is isolated from medicinal plants such as Bergenia crassifolia, Caesalpinia digyna, and Flueggea leucopyrus. Bergenin has been shown to have anti-inflammatory, anti-ulcerogenic, and wound healing properties. Still, its anticancer potential has been realized only recently. We performed a proteomic analysis of cervical carcinoma cells treated with bergenin and found it to influence multiple hallmarks of cancers, including apoptosis, angiogenesis, and tumor suppressor proteins. It was also involved in many different cellular processes unrelated to cancer, as shown by our proteomic analysis. Further analysis showed bergenin to be a potent -angiogenic agent by reducing key angiogenic proteins like Galectin 3 and MMP-9 (Matrix Metalloprotease 9) in cervical carcinoma cells at cellular level as shown by western blotting assay. Further understanding of this interaction was carried out using molecular docking analysis, which indicated MMP-9 has more affinity for Bergenin as compared to Galectin-3. Cumulatively, our data provide novel insight into the anti-angiogenic mechanism of Bergenin in cervical carcinoma cells by modulation of multiple angiogenic proteins like Galectin-3 and MMP-9 which warrant its further development as an anticancer agent in cervical cancer. Citation Format: Mayank Singh, Ravi Chauhan, Lakshay Malhotra, Ashna Gupta, Gunjan Dagar, Dayasagar Das, Shahab Uddin, Ethayathulla Abdul Samath, Muzafar A. Macha, Ammira Al shabeeb Akil, Ajaz A. Bhat. Bergenin inhibits growth of human cervical cancer cells by decreasing galectin-3 and MMP-9 expression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1830.

Growth Inhibition of Standardized Amine Fraction From Clinacanthus nutans on Mice Xenograft Model for Human Cervical Cancer

Introduction: Cervical cancer continue to be a leading cause of cancer-related death for women. The current anticancer drugs remain inefficient due to the lack of specificity to inhibit the cancer cells growth. Plant-based drugs with potent anticancer should add to the efforts in finding drugs with lesser side effects but great potential to increase survival. Hence, this study sought to determine the tumour growth inhibition of standardized fraction (SF1) from Clinacanthus nutans as a potential alternative treatment for cervical cancer. Methods: SF1 was isolated from a series of bioassay-guided fractionation of C.nutans leaves. Female nude mice were xenografted with human cervical cancer cell line, SiHa, subcutaneously. When the tumour volume reached 100mm3, SF1 was intraperitoneally injected once daily for 28 days. Body weight and tumour volume were recorded every 3 days. Tumour and liver were fixed for hematoxylin and eosin staining and immunohistochemistry using caspase-3 antibody. Blood was collected by cardiac puncture for assessment of liver enzymes level, AST and ALT. Results: SF1 has a great tumour inhibition with more than 50% inhibition rate compared to the negative control group (DMSO). ALT and AST levels in SF1-treated mice were maintained in normal ranges compared to the positive control group, cisplatin indicating no sign of toxicities. SF1-treated group revealed no indication of liver toxicity. The number of tumour mitosis was reduced and caspase-3, a critical mediator of apoptosis was overexpressed in SF1-treated group. Conclusion: SF1 demonstrated potent anticancer activity by inhibiting the tumour growth with less toxic effects, suggesting a promising candidate for preventing cancer progression.

Lamin B1 curtails early human papillomavirus infection by safeguarding nuclear compartmentalization and autophagic capacity

Human papillomavirus (HPV) infection is a primary cause of cervical and head-and-neck cancers. The HPV genome enters the nucleus during mitosis when the nuclear envelope disassembles. Given that lamins maintain nuclear integrity during interphase, we asked to what extent their loss would affect early HPV infection. To address this question, we infected human cervical cancer cells and keratinocytes lacking the major lamins with a HPV16 pseudovirus (HP-PsV) encoding an EGFP reporter. We found that a sustained reduction or complete loss of lamin B1 significantly increased HP-PsV infection rate. A corresponding greater nuclear HP-PsV load in LMNB1 knockout cells was directly related to their prolonged mitotic window and extensive nuclear rupture propensity. Despite the increased HP-PsV presence, EGFP transcript levels remained virtually unchanged, indicating an additional defect in protein turnover. Further investigation revealed that LMNB1 knockout led to a substantial decrease in autophagic capacity, possibly linked to the persistent activation of cGAS by cytoplasmic chromatin exposure. Thus, the attrition of lamin B1 increases nuclear perviousness and attenuates autophagic capacity, creating an environment conducive to unrestrained accumulation of HPV capsids. Our identification of lower lamin B1 levels and nuclear BAF foci in the basal epithelial layer of several human cervix samples suggests that this pathway may contribute to an increased individual susceptibility to HPV infection.Graphical abstract

Commercial Wheat and Oat Flour Derived Fluorescent Carbon Quantum Dots through Sustainable Hydrothermal Synthesis and Their Biological Activities

AbstractIn this study, we utilized processed edible commercial wheat and oat flours to synthesize highly fluorescent carbon dots using green hydrothermal synthesis. The as‐synthesized carbon quantum dots (CQDs) were characterized using UV‐vis spectrophotometry, photoluminescence spectrophotometry, high‐resolution transmission electron microscopy, and Fourier transform infrared spectroscopy. The synthesized wheat‐CQDs (W‐CQDs) and oat‐CQDs (O‐CQDs) had monodispersed spherical shapes with average sizes of 3–8 and 5–10 nm, respectively, and exhibited strong fluorescence intensity and excitation‐dependent photoluminescence. The W‐CQDs and O‐CQDs were highly dispersed in aqueous solutions and exhibited bright green and blue fluorescence, respectively, under UV light (λex=380 and 370 nm). Both CQDs contained abundant functional groups, including amino, hydroxyl, and carboxyl groups, which may play a major role in biological applications. The W‐CQDs and O‐CQDs showed good free‐radical scavenging capacity in the 2,2‐diphenylpicrylhydrazyl (DPPH) and H2O2 assays. The anti‐inflammatory activities of W‐CQDs and O‐CQDs were more effective than that of the standard, with EC50 values of 1.8 and 1.3 mg/mL, respectively, in the protein denaturation assay. Additionally, the CQDs exhibited low cytotoxicity toward NIH3T3 fibroblast and human HeLa cervical cancer cells when treated with a 5‐mg/mL concentration. Therefore, green‐synthesized W‐CQDs and O‐CQDs are easily available, less cytotoxic, and applicable to therapeutic and cell nanocarrier purposes.

Engineering MnO2 Nanomulberries for Enhancing PTT/PDT Synergistic Cancer Therapy

AbstractConsidering the great catalyzation ability to degrade H2O2 into O2, MnO2 nanoparticles are utilized as nanocarriers in phototherapies for anticancer treatments to overcome the hypoxia tumor microenvironment (TME). However, the confliction of size requirement between promoting tumor delivery and cellular uptake and catalyzing H2O2 degradation limits the functions of MnO2 nanoparticles. Herein, this work presents strategy to cross‐link small‐sized MnO2 nanoparticles, during which the indocyanine green (ICG) is loaded, thus obtaining large‐sized MnO2 nanomulberries (MnINCs) for overcoming the size requirement confliction. The MnINCs present great performances in both photothermal therapy (PTT) and photodynamic therapy (PDT) effects. This work tests the therapeutic efficacy of MnINCs in various human cancer cell lines including human cervical cancer cell line (HeLa), human breast cancer cell line (MCF‐7), human hepatoma cell line (HepG‐2), human gastric carcinoma cell line (MGC803), and a Hela cell‐derived xenograft model. The satisfying growth inhibition effects are observed in all these cell lines and the tumor model. The demonstrations for using MnINCs to overcome the size requirement confliction between promoting tumor delivery and cellular uptake and catalyzing H2O2 degradation highlight the promising application potentials of the clustering strategy for developing innovative treatments in cancer therapy.

Enhanced photocytotoxicity induced by a triphenylphosphonium-appended platinum diimine complex

Photodynamic therapy (PDT) agents induce photodamage only where photosensitizers localize in tumor cells. Since apoptosis is a major pathway of PDT-mediated cell death in vivo, the therapeutic efficacy of PDT drugs against tumor cells may be improved by regulating intrinsic apoptosis. Mitochondria are the major intracellular organelles that regulate apoptosis, thus triphenylphosphonium, a well-known mitochondria-targeting molecule, was appended to a photoactive platinum diimine complex. The ability of the triphenylphosphonium-appended complex to generate singlet oxygen was assessed by fluorescence spectroscopy using 1,3-diphenylisobenzofuran as a probe. The photocytotoxicity towards HeLa cells (human cervical cancer cell line) or HL-7702 cells (human liver cell line) was assessed by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assays. The platinum complex has moderate capability to generate singlet oxygen and it has more potent photocytotoxicity than its analogue Pt-CA. Both platinum diimine complexes studied are largely nontoxic to human liver cells either in the dark or after irradiation.

In vitro Evaluation of Anti-Cancer Potential of Different Solvent Extracts Derived from Clerodendrum Infortunatum Linn against Cervical Cancer.

Cervical cancer is a prevalent and deadly malignancy in females, with chemotherapy often proving ineffective due to significant side effects and the development of chemo-resistance. This study investigates the medicinal potential of Clerodendrum infortunatum linn. , a genus with approximately 500 species in the Lamiaceae family. Limited research exists on the species of Clerodendrum infortunatum and its various solvent extracts. The study aims to assess the anti-cancer properties of different solvent extracts from this plant on human cervical cancer cells. The study examines the plant's phytochemical components and their potential to inhibit cancer growth. Aerial parts of the plant were extracted using the Soxhlet method, and the presence of Rutin, Quercetin, and Gallic Acid in specific solvent extracts was validated through High-Performance Thin Layer Chromatography (HPTLC). In vitro assays, including MTT, Apoptosis, Cell Cycle analysis, Intracellular Reactive Oxygen Species assessment, and Gene expression PCR, were conducted to investigate the plant's anti-cancer properties further. The outcomes of the phytochemical assessment indicated that Rutin was predominantly present in the water extract, with quercetin being more concentrated in the decoction, and the hydro-alcoholic extract showing elevated levels of gallic acid. Notably, the decoction extract demonstrated the highest cytotoxic activity, primarily through early apoptosis and arrests in the S-phase and G2M phases. Clerodendrum infortunatum exhibited a reduction in Intracellular Reactive Oxygen Species. The gene expression analysis disclosed an impact on the BCL-2 gene. Notably, Clerodendrum infortunatum exhibited the ability to initiate early apoptosis, halt the cell cycle at the S and G2M phases, and diminish levels of reactive oxygen species significantly. The gene expression analysis revealed an influence on the BCL-2 gene. To sum up, this research underscores the encouraging cytotoxic and antioxidant attributes of Clerodendrum infortunatum, implying its potential for cervical cancer treatment.

Qualitative Phytochemical Screening, Fatty Acid Profile and Biological Studies of the Bark of &lt;i&gt;Mallotus nudiflorus&lt;/i&gt; (Pitali) Plant

In the present study, the Mallotus nudiflorus (L.) plant has been taken to determine the in-vitro analysis to find out the therapeutic value. The bioassays of the raw extract of methanol of bark of M. nudiflorus and by Kupchan’s extraction method collecting n-Hexane (HEX), Dichloromethane (DCM), Chloroform (CHCl3), Ethylacetate (EA) and Aqueous (AQ) fractions were scrutinized to find out its therapeutic value. The findings of phytochemical screening of the methanol extract of barks revealed the presence of several secondary metabolites. By using the GC-FID method the result showed that M. nudiflorus contained four bound fatty acids and four free fatty acids. EA fraction had the maximum phenolic content among all the fractions at (133.67±0.99) mg of GAE/g where the Folin- Ciocalteu reagent was used as an oxidizing agent. The antioxidant activity was measured in terms of its ability to scavenge free radicals (DPPH assay). Among all extractives, the greatest ability to scavenge for free radicals was shown by EA extract with an IС50 value (12.08±0.15) μg/ml. In the toxicity of brine shrimp test, the HEX fraction had the maximum toxicity with an LC50 value of (0.12±0.01) μg/ml. Cell cytotoxicity was observed for sample CHCl3 and EA on both the Vero (kidney epithelial cells taken from an African green monkey) and HeLa (a human cervical cancer cell) cell line. All the fractions were subjected to in vitro microbial screening, which revealed that DCM, CHCl3, and EA fractions showed growth inhibition, particularly against various Gram-positive and Gram-negative bacteria by disc diffusion method. The maximum zone of inhibition in the antimicrobial activity was produced by CHCl3 fraction against Staphylococcus aureus (17 mm). The results of in vitro experiments have demonstrated that the extracts from the barks of M. nudiflorus have great potential for medicinal uses and might be studied for further chemical exploration.

Efficient gene delivery by multifunctional star poly (β-amino ester)s into difficult-to-transfect macrophages for M1 polarization

Gene delivery to macrophages holds great promise for cancer immunotherapy. However, traditional gene delivery methods exhibit low transfection efficiency in macrophages. The star-shaped topological structure of polymers is known to encapsulate genes inside their cores, thereby facilitating sustained release of the genetic material. Herein, combining the structural advantages of star polymers and the transfection advantages of poly (β-amino ester)s (PAEs), we developed a novel linear oligomer grafting-onto strategy to synthesize a library of multi-terminal star structured PAEs (SPAEs), and evaluated their gene delivery efficiency in various tissue cells. The transfection with human hepatocellular carcinoma cells (HepG2, HCC-LM3 cells and MHCC-97H cells), rat normal liver cells (BRL-3 A cells), human ovarian cancer cells (A2780 cells), African green monkey kidney cells (Vero cells), human cervical cancer cells (HeLa cells), human chondrosarcoma cells (SW1353 cells), and difficult-to-transfect human epidermal keratinocytes (HaCaT cells) and normal human fibroblast cells (NHF cells) showed that SPAEs exhibited superior transfection profile. The GFP transfection efficiency of top-performing SPAEs in HeLa cells (96.1%) was 2.1-fold, and 3.2-fold higher compared to jetPEI and Lipo3000, respectively, indicating that the star-shaped topological structure can significantly enhance the transfection efficiency of PAEs. More importantly, the top-performing SPAEs could efficiently deliver Nod2 DNA to difficult-to-transfect RAW264.7 macrophages, with a high transfection efficiency of 33.9%, which could promote macrophage M1 polarization and enhanced CD8+ T cell response in co-incubation experiments. This work advances gene therapy by targeting difficult-to-transfect macrophages and remodeling the tumor immune microenvironment.

Downregulation of LAMB3 Altered the Carcinogenic Properties of Human Papillomavirus 16-Positive Cervical Cancer Cells.

Nearly all cervical cancer cases are caused by infection with high-risk human papillomavirus (HR-HPV) types. The mechanism of cervical cell transformation is related to the powerful action of viral oncoproteins and cellular gene alterations. Transcriptomic data from cervical cancer and normal cervical cells were utilized to identify upregulated genes and their associated pathways. The laminin subunit beta-3 (LAMB3) mRNAwas overexpressed in cervical cancer and was chosen for functional analysis. The LAMB3 was predominantly expressed in the extracellular region and the plasma membrane, which play a role in protein binding and cell adhesion molecule binding, leading to cell migration and tissue development. LAMB3 was found to be implicated in the pathway in cancer and the PI3K-AKT signaling pathway. LAMB3 knockdown decreased cell migration, invasion, anchorage-dependent and anchorage-independent cell growth and increased the number of apoptotic cells. These effects were linked to a decrease in protein levels involved in the PI3K-AKT signaling pathway and an increase in p53 protein. This study demonstrated that LAMB3 could promote cervical cancer cell migration, invasion and survival.

Thermally annealed large-scale gold nanostructure platform for long-term and label-free electrochemical monitoring of cellular metabolism

In recent developments, an electrochemical method has emerged as a promising tool that enables real-time and non-destructive monitoring of cell viability and functionality in vitro. However, the restricted cell cultivation area and poor adhesion properties of the gold nanostructure film impede its potential as a new analytical tool for cellular research. In this study, we present an advanced platform designed for prolonged cell growth, differentiation, and drug screening, while providing electrochemical detection capability. The thermal annealing process, implemented after gold nanostructure deposition, was found to induce structural reformation on the indium tin oxide (ITO) film through Ostwald ripening and coalescence, leading to increased surface coverage and reduced height. The resulting optimized platform, referred to as the thermally annealed large-scale gold nanostructure (TLGN), offers an expanded cell culture area (about 227 mm2) with sustained resistance to gold film detachment, a critical challenge observed in previously reported electrodes, lasting up to 4 weeks under exposure to cell culture medium. Using TLGN, redox signals were successfully detected from a substantial number of human cervical cancer cells (470,000 cells) at days in vitro 5, a cell count that is 10.35 times higher compared to our previous gold nanostructure electrode. The cells were continuously monitored for up to 4 days under treatment with the anticancer drug (CB-839). Furthermore, the TLGN platform demonstrated its effectiveness in electrochemically monitoring the osteogenesis of human mesenchymal stem cells over a differentiation period of 28 days, based on observed changes in cellular metabolism. In summary, the TLGN platform proves invaluable not only for assessing drug accumulation and potential cytotoxicity at the cellular level but also for real-time monitoring of mitochondrial dynamics during cellular differentiation.