Project description:We established panels of patient-derived culture of the cancer cells from small cell carcinoma of the cervix uteri (SCCC) by cancer tissue–originated spheroids (CTOS) method. Then we developed in vitro sensitivity assay for radiation using CTOSs to assess the intrinsic radio-sensitivity and mechanism of radio-resistance in individual SCCC patients. To find factors that affect to the radio-sensitivity we compared gene expression of radio-resistant CTOS (cerv-5) and radio-sensitive CTOS (cerv-9).
Project description:We established panels of patient-derived culture of the cancer cells from small cell carcinoma of the cervix uteri (SCCC) by cancer tissueM-bM-^@M-^Soriginated spheroids (CTOS) method. Then we developed in vitro sensitivity assay for radiation using CTOSs to assess the intrinsic radio-sensitivity and mechanism of radio-resistance in individual SCCC patients. To find factors that affect to the radio-sensitivity we compared gene expression of radio-resistant CTOS (cerv-5) and radio-sensitive CTOS (cerv-9). We compared gene expression of cerv5 and cerv9 CTOSs under the culture condition.
Project description:Expression profiling of nasopharyngeal carcinoma patients comparing radio-sensitive samples with radio-resistant samples. Two condition-experiments, radio-sensitive and radio-resistant nasopharyngeal carcinoma patients. Biological replicates: 8 radio-sensitive, 12 radio-resistant, different donors in the same hospital. One patient per array.
Project description:Small cell neuroendocrine carcinoma (SCNEC) of the uterine cervix is a rare disease with a poor prognosis. The lack of established disease models has hampered therapy development. We established a panel of organoid from patients of SCNEC of the uterine cervix. Sensitivity assays against clinically used drugs revealed remarkable variations between the lines, and we successfully identified specific gene sets which likely contribute to the sensitivity to the tested drugs. Of note, we found one line which was exceptionally sensitive to irinotecan, and investigated the mode of action in the case. Chemotherapeutic drug sensitivity assays using SCNEC CTOS lines obtained from primary patient samples with a high success rate may provide useful information for treating SCNEC.
Project description:Mitogen activated protein kinase (MAPK) inhibitors are important therapies for treating many cancers. However, the development of acquired resistance to most protein kinase inhibitors limit their ability to provide durable responses. Approximately 50% of malignant melanomas contain activating mutations in the BRAF kinase, which promote cancer cell survival by activating the extracellular signal-regulated kinase-1/2 (ERK1/2) pathway through direct phosphorylation of the MAPK/ERK kinase-1/2 (MEK1/2). Although combination treatment with BRAF and MEK1/2 inhibitors is a recommended approach to treat melanoma, the development of drug resistance remains a barrier to achieving long term patient benefits. Using high-resolution label-free mass spectrometry, the current studies compared relative protein changes in BRAF and MEK1/2 inhibitor resistant A375 melanoma cells grown as monolayers or 3D spheroids. While approximately 66% of proteins identified were common in both monolayer and spheroid cultures, only 6.2% or 3.6% of proteins that significantly increased or decreased, respectively, compared to drug sensitive cells were common between the drug-resistant monolayer and spheroid cells. Major changes in drug-resistant monolayers suggested upregulation of alternative kinase signaling pathways that promote growth and metastasis. In contrast, the major changes in drug-resistant spheroids indicated increased catabolic metabolism to support oxidative phosphorylation and energy requirements.
Project description:Cisplatin resistance is a problem in cancer treatment. Using DNA microarray, we detected differentially expressed genes in cisplatin-resistant cervix carcinoma HeLa cells compared to parental cells.
Project description:Small cell neuroendocrine carcinoma of the cervix (SCNEC) is classified as a high-grade neuroendocrine carcinoma of the cervix and has a worse prognosis than other major histological types of cervical cancer, such as squamous cell carcinoma and adenocarcinoma, due to the lack of effective treatments. Developing novel therapeutic targets based on its molecular characteristics is highly desirable; however, it has been limited due to the rarity of SCNEC and the resulting lack of research resources. Herein, we identified vaccinia-related kinase 1 (VRK1) as a potential therapeutic target for SCNEC. VRK1 was focused via our previous reported proteomic analysis of patient-derived organoids. Immunohistochemistry of patient samples revealed that VRK1 expression is consistently high in SCNEC, whereas its expression is variable in other cervical carcinomas. VRK1 knockdown significantly suppressed cell proliferation in three-dimensional cultures and xenograft tumor growth in vivo. Gene Set Enrichment Analysis of RNA-seq data obtained from mouse xenograft models indicated that VRK1 is associated with mitochondrial function. Subsequent in vitro experiments demonstrated that VRK1 knockdown suppressed SCNEC cell proliferation and simultaneously impaired mitochondrial function under the unique condition with oxidative stress. Finally, low VRK2 expression, proposed as a biomarker for the effectiveness of VRK1 knockdown, was consistently observed in SCNEC. These findings suggest that SCNEC is an optimal tumor group for targeting VRK1. VRK1 knockdown induced significant anti-tumor effects, highlighting its potential as a therapeutic target for SCNEC. Mechanistically, these effects appear to be mediated through mitochondrial dysfunction.
