Breast cancer is a major cause of cancer-related death among women. Tumor protein D52-like 2 (TPD52L2) is one member of the TPD52 family, which has been shown to function in mediating cell proliferation, apoptosis, and vehicle trafficking. TPD52 was originally identified in human breast carcinoma. In this study, the authors found that TPD52L2 is extensively expressed in multiple human breast cancer cell lines. To elucidate the functional role of TPD52L2 in breast cancer, the authors employed lentivirus-mediated short hairpin RNA (shRNA) to knock down TPD52L2 in one breast cancer cell line, ZR-75-30, which showed high TPD52L2 expression. The shRNA-mediated TPD52L2 knockdown inhibited the proliferation and colony formation in ZR-75-30 cells, as determined by MTT and colony formation assays. Knockdown of TPD52L2 led to an accumulation of cells in the G0/G1 phase of the cell cycle. Furthermore, knockdown of TPD52L2 promoted GSK3β phosphorylation in ZR-75-30 cells. This investigation indicates that TPD52L2 plays an essential role in the growth of breast cancer cells, which may contribute to provide gene therapy for breast cancer treatment.
Summary form only given. Breast cancer is one of the major malignant tumors for women, which has the highest incidence for women in America, and cause the second highest mortality among other cancers. Traditional treatment of breast cancer includes surgery, radiation treatment and chemotherapy, most of which have various disadvantages and side effects. Gemcitabine is a clinical used chemotherapeutic drug, inhibiting tumor through cytotoxic activity .In this study, a novel promising technique in cancer therapy, the nanosecond pulsed electric fields (known as nsPEFs), was applied in combination with Gemcitabine to the treatment of 2 types of human breast cancer cell lines MCF-7 and MDA MB-231. Both kinds of cells were exposed to nsPEFs before treated with Gemcitabine. Significant inhibitory effects were achieved in both cell lines, verified by MTT tests (viability test). A 5-fold increase of cell inhibition was detected in cells pulsed before exposing to Gemcitabine compared to treated by Gemcitabine alone. The typical morphological changes of cell apoptosis were observed by TEM, and apoptosis evaluated by cytoflowmetry. Transwell invasion assay and clonogenic assay indicated a decrease in the cell ability of migration and proliferation after the combined treatment. It is hypothesized that nsPEFs triggered cell functions, inducing cell apoptosis, and enhanced the inhibition effects in the treatment of Gemcitabine after nsPEFs exposure. This novel combined approach in breast cancer treatment may have potential applications in clinical medicine.
Background: Messenger RNA (mRNA)-based immunogene therapy holds significant promise as an emerging tumor therapy approach. However, the delivery efficiency of existing mRNA methods and their effectiveness in stimulating anti-tumor immune responses require further enhancement. Tumor cell lysates containing tumor-specific antigens and biomarkers can trigger a stronger immune response to tumors. In addition, strategies involving multiple gene therapies offer potential optimization paths for tumor gene treatments. Methods: Based on the previously developed ideal mRNA delivery system called DOTAP-mPEG-PCL (DMP), which was formed through the self-assembly of 1.2-dioleoyl-3-trimethylammonium-propane (DOTAP) and methoxypoly (ethylene glycol)-b-poly (ϵ-caprolactone) (mPEG-PCL), we introduced a fused cell-penetrating peptide (fCPP) into the framework and encapsulated tumor cell lysates to form a novel nanovector, termed CLSV system (CLS: CT26 tumor cell lysate, V: nanovector). This system served a dual purpose of facilitating the delivery of two mRNAs and enhancing tumor immunogene therapy through tumor cell lysates. Results: The synthesized CLSV system had an average size of 241.17 nm and a potential of 39.53 mV. The CLSV system could not only encapsulate tumor cell lysates, but also deliver two mRNAs to tumor cells simultaneously, with a transfection efficiency of up to 60%. The CLSV system effectively activated the immune system such as dendritic cells to mature and activate, leading to an anti-tumor immune response. By loading Bim-encoded mRNA and IL-23A-encoded mRNA, CLSV/Bim and CLSV/IL-23A complexes were formed, respectively, to further induce apoptosis and anti-tumor immunity. The prepared CLSV/dual-mRNA complex showed significant anti-cancer effects in multiple CT26 mouse models. Conclusion: Our results suggest that the prepared CLSV system is an ideal delivery system for dual-mRNA immunogene therapy. Keywords: mRNA gene therapy, dual-mRNA delivery, tumor cell lysates, Bim and IL-23A mRNAs, immune response
Chemotherapy either before or after surgery is a common breast cancer treatment. Long-term, high dose treatments with chemotherapeutic drugs often result in undesirable side effects, frequent recurrences and resistances to therapy.The anti-cancer drug, gemcitabine (GEM) was used in combination with pulse power technology with nanosecond pulsed electric fields (nsPEFs) for treatment of human breast cancer cells in vitro. Two strategies include sensitizing mammary tumor cells with GEM before nsPEF treatment or sensitizing cells with nsPEFs before GEM treatment. Breast cancer cell lines MCF-7 and MDA-MB-231 were treated with 250 65 ns-duration pulses and electric fields of 15, 20 or 25 kV/cm before or after treatment with 0.38 μM GEM.Both cell lines exhibited robust synergism for loss of cell viability 24 h and 48 h after treatment; treatment with GEM before nsPEFs was the preferred order. In clonogenic assays, only MDA-MB-231 cells showed synergism; again GEM before nsPEFs was the preferred order. In apoptosis/necrosis assays with Annexin-V-FITC/propidium iodide 2 h after treatment, both cell lines exhibited apoptosis as a major cell death mechanism, but only MDA-MB-231 cells exhibited modest synergism. However, unlike viability assays, nsPEF treatment before GEM was preferred. MDA-MB-231 cells exhibited much greater levels of necrosis then in MCF-7 cells, which were very low. Synergy was robust and greater when nsPEF treatment was before GEM.Combination treatments with low GEM concentrations and modest nsPEFs provide enhanced cytotoxicity in two breast cancer cell lines. The treatment order is flexible, although long-term survival and short-term cell death analyses indicated different treatment order preferences. Based on synergism, apoptosis mechanisms for both agents were more similar in MCF-7 than in MDA-MB-231 cells. In contrast, necrosis mechanisms for the two agents were distinctly different in MDA-MB-231, but too low to reliably evaluate in MCF-7 cells. While disease mechanisms in the two cell lines are different based on the differential synergistic response to treatments, combination treatment with GEM and nsPEFs should provide an advantageous therapy for breast cancer ablation in vivo.
Objective: Glioma is the most common and fatal primary brain tumor that has a high risk of recurrence in adults. Identification of predictive biomarkers is necessary to optimize therapeutic strategies. This study investigated the predictive efficacy of a previously identified radiosensitivity signature as well as Exportin 1 (XPO1) expression levels. Methods: A total of 1,552 patients diagnosed with glioma were analyzed using the Chinese Glioma Genome Atlas and The Cancer Genome Atlas databases. The radiosensitive and radioresistant groups were identified based on a radiosensitivity signature. Patients were also stratified into XPO1-high and XPO1-low groups based on XPO1 mRNA expression levels. Overall survival rates were compared across patient groups. Differential gene expression was detected and analyzed through pathway enrichment and Gene Set Enrichment Analysis (GSEA). To predict 1-, 3-, and 5-years survival rates for glioma patients, a nomogram was established combining the radiosensitivity gene signature, XPO1 status, and clinical characteristics. An artificial intelligence clustering system and a survival prediction system of glioma were developed to predict individual risk. Results: This proposed classification based on a radiosensitivity gene signature and XPO1 expression levels provides an independent prognostic factor for glioma. The RR-XPO1-high group shows a poor prognosis and may benefit most from radiotherapy-combined anti-XPO1 treatment. The nomogram based on the radiosensitivity gene signature, XPO1 expression, and clinical characteristics performs more optimally compared to the WHO classification and IDH status in predicting survival rates for glioma patients. The online clustering and prediction systems make it accessible to predict risk and optimize treatment for a special patient. The cell cycle, p53, and focal adhesion pathways are associated with more invasive glioma cases. Conclusion: Combining the radiosensitivity signature and XPO1 expression is a favorable approach to predict outcomes as well as determine optimal therapeutic strategies for glioma patients.
Oral squamous cell carcinoma (OSCC) is the most widespread malignant oral cavity neoplasm, with even higher proportion of deaths than breast cancer and cervical cancer. Although advances were made in conventional treatment for OSCC such as surgery and radiation therapy, there has not been significant increase in the 5-year survival rate in the past four decades, and chemotherapeutic drug resistance and side-effects are still main obstacles for chemotherapy. Thus, improvement in the treatment of OSCC is strongly expected. In this study, nanosecond pulsed electric fields, known as nsPEFs, were applied in the treatment of OSCC in combination with gemcitabine, a clinical-used anti-cancer drug. Synergistic effects of nsPEFs and gemcitabine were observed and investigated. Cal-27 cells were exposed to 20 pulses with duration of 100ns at electric field intensity of 10, 30 and 60 kV/cm, and then cultured in the medium with 0.01ug/mL gemcitabine. Cell inhibition was increased by 5 fold after the combined treatment compared to gemcitabine or nsPEFs alone, verified with MTT test, and remarkable synergistic effects were obtained with synergistic quotient over 3. Cell proliferation and invasion ability decreased significantly, determined by clonogenic assay and transwell invasion assay. Cell apoptosis was evaluated by flow cytometry and morphological changes were observed by TEM. It is indicated that the combined treatment has significant advances over either gemcitabine or nsPEFs, and synergistic inhibition in cell line cal-27 was demonstrated. The combined therapy may reduce the dose of both chemotherapeutic drug and nsPEF, and thus may avoid side-effects or drug resistances as well as provide a better treatment for oral cancer.
Application of multiphoton microscopy (MPM) to clinical cancer research has greatly developed over the last few years. In this paper, we mainly focus on two-photon excitation fluorescence (TPEF) and second harmonic generation (SHG) for investigating esophageal cancer. We chiefly discuss the SHG/TPEF image and spectral characteristics of normal and cancerous esophagus submucosa with the combined multi-channel imaging mode and Lambda mode of a multiphoton microscope (LSM 510 META). Great differences can be detected, such as collagen content and morphology, glandular-shaped cancer cells, TPEF/SHG intensity ratio, and so on, which demonstrate that the multiphoton imaging technique has the potential ability for minimally-invasive early cancer diagnosis.
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