ABSTRACT
OBJECTIVE: To use microarray chip technology for screening of stem cell radiation related miRNAs in laryngeal squamous cell carcinoma; study and explore the relationship of miRNAs with radiosensitivity of laryngeal squamous cells. METHOD: After conventional culture and amplification of the laryngeal squamous carcinoma cell line Hep-2, CD 133+ cells were screened out with combination of isolated culture of stem cell microspheres and FACS for preparation of laryngeal cancer stem cells. After radiation treatment, miRNAs of laryngeal squamous carcinoma stem cells before and after radiation were enriched and purified. After microarray hybridization with mammalian miRNA and scanning of fluorescence signal, the miRNAs of laryngeal squamous carcinoma stem cells before and after radiation was subject to differential screening and clustering analysis. Real-time quantitative RT-PCR was used to verify part of the differentially expressed miRNAs. RESULTS: 70 miRNAs related to laryngeal cancer stem cell radiation with 2-fold difference in expression were screened out, in which 62 were down-regulated and 8 were up-regulated. Fluorescent quantitative RT-PCR results were consistent with miRNAs chip results. CONCLUSION: Some miRNAs may be involved in self-regulation with laryngeal squamous carcinoma stem cell radiation.
Subject(s)
Biomarkers, Tumor/genetics , Carcinoma, Squamous Cell/genetics , Gene Expression Profiling , Laryngeal Neoplasms/genetics , MicroRNAs/genetics , Neoplastic Stem Cells/radiation effects , Radiation Tolerance/genetics , Animals , Carcinoma, Squamous Cell/radiotherapy , Humans , Laryngeal Neoplasms/radiotherapy , Neoplastic Stem Cells/metabolism , Oligonucleotide Array Sequence Analysis , RNA, Messenger/genetics , Real-Time Polymerase Chain Reaction , Reverse Transcriptase Polymerase Chain Reaction , Tumor Cells, Cultured , X-RaysABSTRACT
OBJECTIVE: To examine the influence of chemotherapy with FOLFOX protocol (CT-F) on the immunologic function in patients with advanced colorectal cancer. METHODS: A total of 43 patients with advanced colorectal cancer were included. Patients who received FOLFOX chemotherapy (Group A, n=22) were compared to those who did not(Group B, n=21). Blood was obtained from peripheral vein before chemotherapy (T0), at the time of completion of chemotherapy (T1), and 3 months after completion of chemotherapy (T2) to detect the percentage of regulatory T lymphocytes (CD4+CD25+Foxp3+T cells) in total T lymphocytes using fluorescence activated cell sorter (FACS). The level of Th1/Th2 cytokines in the serum including interleukin-2 (IL-2), interleukin-4 (IL-4), interleukin-10 (IL-10) and interferon-γ (IFN-γ) were detected by enzyme-linked immunoabsorbent assay(ELISA). RESULTS: The percentage of regulative T lymphocyte was significantly lower at T1, which increased after chemotherapy but was still lower than that at T0 and that in Group B [(4.15±0.56)%, (5.60±0.88)%, and(5.38±0.92)%, all P<0.01]. The levels of IL-4, IL-10 decreased at T1, and increased to the normal level at T2 compared with those at T0 or those in the control group (all P<0.01). In contrast, the levels of IL-2 and IFN-γ increased significantly at T1 and decreased to the normal level at T2 during the entire observation period. CONCLUSION: For patients with advanced colorectal cancer, FOLFOX chemotherapy can decrease the proportion of regulatory T lymphocytes and results in Th1/Th2 cytokine drift to Th1 type. Therefore, FOLFOX may help the release from the anticancer immune inhibition.