Cell Journal最新文献

Objective: Colorectal cancer (CRC) is the second leading cause of cancer-related deaths worldwide. Engineered biomolecules can be used as a targeted tool to deliver drugs directly to tumors that reduce the adverse effects of conventional treatments. We aimed to prepare non-targeted oxaliplatin-loaded chitosan nanoparticles (OXPT-CS NPs) and targeted OXPT-CS NPs decorated with cetuximab single-chain variable fragment (scFv) to send both NPs to epidermal growth factor receptor (EGFR) overexpressing HCT 116 cells, a human colorectal carcinoma cell line, for comparing their cytotoxicity.

Materials and methods: In this experimental study, OXPT-CS NPs were synthesized using a fluid system. Encapsulation efficiency percentage (EE%) and oxaliplatin release rate were evaluated. Western blot and cell-based ELISA confirmed scFv production and its binding ability to EGFR, respectively. The Fourier transform infrared spectroscopy (FTIR) determined the conjugation of scFv to OXPT-CS NPs. The NPs were characterized, and their toxicity against the HCT 116 cells was evaluated using 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) and flow cytometry assays.

Results: The EE% of OXPT-CS NPs was 93%, and the average diameters were 75.85 ± 8.81 nm and 92.48 ± 9.51 before and after scFv conjugation, respectively. The scFv was purified via affinity chromatography. The western blot method and cell-based ELISA revealed successful purification of scFv and its attachment to EGFR on HCT 116 cells. The FTIR analysis determined the interactions between the scFv and OXPT-CS NPs. According to MTT and flow cytometry results, the targeted delivery system significantly reduced HCT 116 cancer cell viability and increased apoptosis induction up to 99.8%.

Conclusion: The scFv-OXPT-CS NPs demonstrated an increased cytotoxic function due to the presence of scFv in its formulation. This delivery system offers a promising method for delivering chemotherapy drugs to cancer cells. More research is needed on the best strategies for improving treatment efficacy by targeting cancer cells.

Cell-based therapy has shown promising outcomes in the treatment of cerebral palsy (CP). However, there is no consensus on a standard therapeutic protocol regarding the source of cells, optimal cell dose, timing and frequency of cell injections, route of administration, or the use of combination therapy. This lack of consensus necessitates a comprehensive investigation to clarify these crucial yet undefined factors in cell-based therapy for CP patients. In this commentary, we discuss and compare the trends in Gross Motor Function Measure-66 following intrathecal injection of umbilical cord blood mononuclear cells (UCB-MNCs) and umbilical cord tissue mesenchymal stromal cells (UCTMSCs) in children with CP. Our study revealed that MNC injections led to earlier improvements in gross motor function, whereas MSC applications resulted in more sustainable changes. These findings provide key insights into the efficacy of different cell types, which will be beneficial for future studies and for refining cell-based therapy protocols for CP treatment.

Objective: Colorectal cancer (CRC) is the third most common cancer and the second leading cause of cancerrelated deaths worldwide, posing a significant public health challenge. Recent advances in molecular research and technologies have highlighted the potential of long non-coding RNAs (lncRNAs) as key players in CRC development and progression. High-throughput technologies, such as RNA sequencing (RNA-seq), enable comprehensive analysis of cancer omics data, providing valuable insights into CRC biology. This study aimed to identify and validate lncRNAs with potential roles in CRC pathogenesis.

Materials and methods: In this experimental study, we conducted RNA-seq analysis using data from The Cancer Genome Atlas (TCGA) to identify differentially expressed lncRNAs between 481 CRC and 41 healthy control samples. Using the Boruta feature selection algorithm, we identified statistically significant lncRNAs. Based on a comprehensive literature review, four lncRNAs (LINC01730, LINC02487, LINC01836, and LINC01594) were selected for experimental validation. Real-time reverse transcription polymerase chain reaction (RT-PCR) was performed on 10 primary CRC and 9 normal colorectal tissue samples to quantify the expression of these lncRNAs. Additionally, bioinformatic analyses were conducted to explore molecular pathways associated with these lncRNAs in CRC.

Results: RNA-seq analysis identified 388 differentially expressed lncRNAs in CRC tissues compared to healthy controls. Among these, four lncRNAs (LINC01730, LINC02487, LINC01836, and LINC01594) were validated through real-time RT-PCR, confirming their significant upregulation in CRC tissues. Bioinformatic analysis revealed their potential involvement in molecular pathways critical to tumor growth and metastasis.

Conclusion: This study identified and validated four significantly upregulated lncRNAs (LINC01730, LINC02487, LINC01836, and LINC01594) in CRC tissues, providing evidence of their potential roles in CRC pathogenesis. These lncRNAs could serve as promising candidates for future biomarker development and therapeutic research in colorectal cancer.

Objective: Polycystic ovary syndrome (PCOS) is one of the most important causes of infertility, irregular menstrual cycles, and anovulation in women. The current study aimed to investigate the therapeutic effects of Althaea officinalis L. (A. officinale) extract on PCOS in rats.

Materials and methods: In this experimental study, 70 rats in 7 groups (n=10/group) were studied for three weeks as follows; healthy control (HC), patient (PCOS), metformin (PCOS+MET), A. officinale treatment (PCOS+250 and 500 mg/kg A. officinale) and synergistic (PCOS+MET+250 and 500 mg/kg A. officinale) groups. Luteinizing hormone (LH), folliclestimulating hormone (FSH), progesterone (P) and testosterone (T) levels as well as inflammatory cytokines were measured. Total antioxidant capacity and lipid peroxidation levels were analyzed in ovarian tissue. The expression of GLUT-4, AKT, PI3K, PTEN genes and Ki-67 was assessed by real-time polymerase chain reaction (PCR) and immunohistochemistry, respectively.

Results: A. officinale alone and especially in combination with MET moderated inflammatory and antioxidant parameters compared to the PCOS and MET groups. A. officinale in synergistic groups increased the apoptosis of granulosa cells by activating the PI3K/AKT pathway, resulting in a rise in the number of Ki-67 positive cells (P<0.05). Furthermore, following A. officinale treatment the LH/FSH rate decreased and FSH and P increased (P<0.05). Also, A. officinale extract could effectively normalize estrus cycle duration close to the normal group.

Conclusion: The extract of A. officinale, in combination with metformin, can enhance the hypothalamic-pituitaryovary (HPO) axis with synergistic anti-inflammatory and antioxidant effects. Additionally, the extract showed apoptotic effect on cystic granulosa cells.

Veno-occlusive disease with immunodeficiency (VODI) syndrome is a rare genetic disorder characterized by immune system irregularities and a significant mortality rate, despite its infrequency. SP110, situated on chromosome 2q37.1, plays a pivotal role in VODI syndrome, and its association with tuberculosis has been extensively studied. The identification of SP110 mutations holds promise for accelerating the diagnosis and treatment of VODI syndrome, by providing a comprehensive panel for diagnosis and potentially leading to targeted therapies. In this case study, we examined a three-year-old girl born to a consanguineous union who was suspected of having an immunodeficiency disorder. Whole-exome sequencing (WES) and clinical assessments were conducted to screen for and confirm potentially pathogenic mutations. The detected mutation was further analyzed using bioinformatics tools to forecast its impact on protein structure. WES analysis revealed a novel deletion-insertion mutation, c.1181-1182delAGinsT, within SP110. Protein analysis indicated substantial structural modifications in the SP110 protein. This study identified a novel deletion-insertion mutation as a potential contributor to VODI syndrome by affecting the functionality of the SP110 protein. By including various mutations associated with the SP110 gene, this study aimed to expedite diagnosis by creating a comprehensive panel for VODI syndrome.

Objective: CD22, as a surface protein of B cells, is used in the diagnosis and target-specific immunotherapy of B-cell malignancies. SpyTag and SpyCatcher, on the other hand, are two covalently coupled proteins capable of developing a bi- or multi-specific modular protein. The aim of this study was to develop FITC-conjugated SpyCatcher-SpyTagged anti-CD22 Nanobody (FITC-SpyC-SpyT-CD22Nb) to recognize CD22 on the surface of malignant B cells.

Materials and methods: In this experimental study, the SpyTag-CD22Nb construct was subcloned into a pET22 vector and expressed in E. coli BL21 (DE3). After purification using His-tag affinity chromatography, the size of the eluted protein was confirmed on a Western blot. In addition, the SpyCatcher protein, subcloned into pET28, was expressed in E. coli BL21 (DE3), purified by His-tag affinity chromatography and subjected to FITC labeling. FITC-SpyCatcher and SpyTag-CD22Nb were coupled in a 1:1 molar ratio. The specific binding of the produced FITC-SpyC-SpyT-CD22Nb was tested using CD22+ Raji and CD22- K562 cell lines and was evaluated by flow cytometry.

Results: SpyTag-CD22Nb and SpyCatcher were successfully expressed in E. coli BL21 (DE3). The 1:1 molar ratio of SpyTag-CD22Nb and FITC-SpyCatcher successfully formed FITC-SpyC-SpyT-CD22Nb at room temperature. The flow cytometry results showed that FITC-SpyC-SpyT-CD22Nb specifically binds to the CD22+ Raji cells, while there is no binding to the CD22- K562 control cells.

Conclusion: The novel FITC-SpyC-SpyT-CD22Nb produced in the present study is capable of detecting the surficial expression of CD22. According to our findings, FITC-SpyC-SpyT-CD22Nb is applicable for specific targeting of CD22 in a therapeutic manner, i.e., chimeric antigen receptor (CAR)-T cell therapy and antibody drug conjugates (ADCs).

Objective: The spliced form of X-box binding protein 1 (XBP1s) is a key transcription factor in the unfolded protein response (UPR), an adaptive mechanism for cell survival. Many studies demonstrated the induced expression of XBP1s in various cancers, including hepatocellular carcinoma (HCC). Such upregulated expression is linked to an enhancement of cell proliferation, migration, and improvement of the survival rate. In this study, we aimed to assess the therapeutic potential of targeting XBP1s, by specific decoy oligodeoxynucleotide (ODN) and evaluated the cancerous phenotypes in Huh-7 cells.

Materials and methods: In this experimental study, we transfected Huh-7 cells with XBP1s decoy oligonucleotide (ODN). Subsequently, we assess some cellular features, including viability, migration capacity, proliferation potential, and apoptosis. Therefore, various techniques included wound healing test, BrdU, and annexin/PI assays. Additionally, the colony formation capacity was evaluated. The mRNA expression levels of BAX, BCL-2, c-MYC, CCND1, MMP-9, CDH1, and CD133 were quantified by the reverse transcription-quantitative polymerase chain reaction (RT-qPCR).

Results: Transfection of Huh-7 cells by XBP1s decoy ODN led to significant down-regulation of c-Myc, CCND1, MMP-9, BCL-2 and CD133 and up-regulation of CDH1 and BAX transcriptional expressions in comparison with the vehicle group. Our results also demonstrated that transfection of XBP1s-decoy reduced HCC cell viability, proliferation, migration capacity as well as colonization ability in comparison with the vehicle group.

Conclusion: These findings proposed the potential application of XBP1s-decoy ODN to reduce cancerous phenotypes such as cell proliferation, cell migration and apoptosis induction in the Huh-7 cell line. More experiments on other cell lines and primary cells could validate our results.

Objective: Breast cancer is a prevalent and heterogeneous disease, with human epidermal growth factor receptor-2 (HER2) overexpression occurring in over 20% of cases. Poncirin, a biologically active flavonone derived from the immature dried fruits of Poncirus trifoliata, is a 7-O-neohesperidoside of isosakuranetin with a well-documented history in traditional Chinese medicine for its health-promoting properties. While the previous research hinted at its potential as an anticancer agent, its specific effects on HER2 overexpressing breast cancer cells remain largely unexplored. The aim of this study is to investigate the specific effects of Poncirin, on HER2 overexpressing breast cancer cells.

Materials and methods: In experimental study, we assessed cell proliferation using the CCK-8 assay and explored cell migration and invasion with transwell assays. Additionally, we evaluated colony formation ability and examined apoptosis through the acridine orange/ethidium bromide (AO/EB) and Annexin V-fluorescein isothiocyanate (FITC)/ propidium iodide (PI) staining methods. The study also delved into the molecular mechanisms involved by scrutinizing the phosphatidylinositol 3-kinase/serine-threonine protein kinase (PI3K/AKT) signaling pathway via Western blotting. Furthermore, the researchers conducted in vivo experiments using mouse models to corroborate the findings in a living organism.

Results: Poncirin demonstrated a remarkable ability to selectively inhibit proliferation and metastasis of HER2 overexpressing breast cancer cells. Mechanistically, the compound seemed to exert its effects by modulating the PI3K/AKT signaling pathway, implying its central role in the observed anticancer effects. These findings were further substantiated by in vivo experiments, which consistently showed a reduction in tumor growth when poncirin was administered.

Conclusion: This study underscores potential of poncirin as a potent agent for restraining the growth and metastasis of HER2 overexpressing breast cancer cells. The evidence suggests that poncirin efficacy may be attributed to its modulation possibly through PI3K/AKT pathway.

Given the critical role of human papillomavirus (HPV) in the cause of cervical cancer and other malignancies, there is a need for innovative approaches to preventing this infection. It has been shown that immunoinformatics is an important strategy in computational vaccinology. It is used to design new multi-epitope vaccines against different types of HPV and subsequent cervical cancer. This paper reviews the scope of the entire computational pipeline of HPV vaccine design, starting from data analysis at the genomic and proteomic levels and continuing to epitope predictions of the innate and adaptive immune systems. The search strategy was based on investigating original articles published in "Google Scholar" and "PubMed" from 2015 to 2023-2024. The terms "Immunoinformatics", "Bioinformatics", "Human papillomavirus (HPV)", "Vaccine design", "In silico vaccine design", "Multi-epitope vaccine design", "Vaccinology" and "HPV vaccine" were used to for this purpose. We discussed various essential tools involved in the computational design of the vaccine process, e.g., sequence analysis, epitope prediction, conservancy analysis, tertiary structure modeling, refinement, molecular docking, molecular dynamics (MD) simulation, and in silico cloning. This review article describes immunoinformatics methods that facilitate the design of a multi-epitope vaccine against HPV. However, this pipeline can also be used to design novel chimeric vaccines for other pathogens.

Objective: Head and neck squamous cell carcinoma (HNSCC) with a high mortality rate is among the most common types of cancer in the world. Human epidermal growth factor receptor 2 (HER2) is expressed higher than normal level in the most HNSCC tumors, making them resistant to chemotherapy and radiotherapy. Therefore, HER2 has been introduced as a suitable target for anticancer drugs. The aim of this study is to examine the efficacy of a treatment protocol involving targeted delivery of idarubicin encapsulated in trastuzumab-decorated liposomes to HNSCC cells.

Materials and methods: In the current experimental study, efficacies of idarubicin, prepared liposomal idarubicin, and constructed immunoliposomal idarubicin (trastuzumab-decorated) were investigated in killing HN5 cells, a HER2- overexpressing HNSCC-originating cell line. Liposomal content of idarubicin and trastuzumab were qualified by UVVisible spectroscopy and preparations were characterized for shape and size by atomic force microscopy (AFM) and dynamic light scattering (DLS). To clarify role of the missing parts of the available crystal structure (PDB ID: 1n8z) within trastuzumab-HER2 interactions, we used a 40 ns molecular dynamic simulation approach.

Results: Based on the obtained results, liposomal idarubicin showed higher toxicity of the encapsulated drug on HN5 cells compared to the traditional free drug formulations. The immunoliposomal form of idarubicin was more effective than the liposomal formulation, in killing of HN5 cells. In addition, simulation of interactions between trastuzumab and HER2 revealed that the missing parts (in the crystal structure) of HER2 have critical interaction with trastuzumab, through salt-bridges and hydrogen bonds.

Conclusion: It seems that the prepared immunoliposomes could attach more efficiently to HER2 overexpressing cells, which consequently leads to increasing cellular uptake of idarubicin through a receptor-mediated endocytosis mechanism. Moreover, simulation of the interaction between HER2 and trastuzumab suggested considerable possibilities for increasing trastuzumab affinity to HER2.