ABSTRACT
Objective To investigate the effect of miR-1269a expression on the radiotherapy sensitivity of H460 stem cell-like cells of non-small cell lung cancer. Methods Non-small cell lung cancer H460 cell line was selected for serum-free suspension culture, and CD133 and CD44 positive H460 stem cells were screened by flow cytometry. qRT-PCR was performed to detect the expression levels of miR-30a, miR-148a, miR-148b, miR-152, miR-411, miR-497, miR-598, miR-424, miR-761, miR-1269a, miR-1280 and CD44, CD133 mRNA in H460 stem cell-like cells. The effects of miR-1269a expression on the radiotherapy sensitivity of H460 stem cell-like cells were analyzed using CCK-8 experiments, stem cell pelleting experiments, flow cytometry, and comet experiments. Results The expressions of CD133 and CD44 mRNA increased 1.30±0.03 times and 1.19±0.02 times, respectively, in H460 stem cell-like cells than in H460 cells (P<0.05). The expression level of miR-1269a was the highest in H460 stem cell-like cells, and was higher than that in H460 cells (P<0.05). After transfection of miR-1269a inhibitor and X-ray treatment, the growth inhibition rate of H460 stem cell-like cells increased significantly (72.11%±2.45%), the apoptosis rate reached to 17.70%±0.54%, and the fragmentation of DNA double-strand breaks in cells also increased significantly. After 2 Gy of X-ray radiation, the tail moment was 1.19±0.02 times of that before transfection, implying that transfection of miR-1269a inhibitor significantly increased the sensitivity of H460 stem cell-like cells to radiotherapy (P<0.05). After transfection of pcDNA3.1-SOX7, the effect of X-ray treatment on H460 stem cell-like cells was similar to that of miR-1269a inhibitor transfection. Simultaneous transfection of pcDNA3.1-SOX7 and miR-1269a mimics reversed the inhibitory effect of transfection of pcDNA3.1-SOX7 on the malignant biological behavior of H460 stem cell-like cells. Conclusion Inhibition of miR-1269a may enhance the radiotherapy sensitivity of non-small cell lung cancer, and the mechanism may be caused by up-regulation of SOX7 expression.
ABSTRACT
<p><b>OBJECTIVE</b>To examine the influence of chemotherapy with FOLFOX protocol (CT-F) on the immunologic function in patients with advanced colorectal cancer.</p><p><b>METHODS</b>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).</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>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.</p>