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. 2022 Aug 24;10(8):e004832. doi: 10.1136/jitc-2022-004832

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© Author(s) (or their employer(s)) 2022. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.

This is an open access article distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited, appropriate credit is given, any changes made indicated, and the use is non-commercial. See http://creativecommons.org/licenses/by-nc/4.0/.

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Depletion of mir-21 potentiates response to chemotherapy in oral cancer cells. (A) Left, genomic sequence of mir21a-5p of wild-type murine oral cancer cell line MTCQ1 (MTCQ-WT) and mir-21 knockout subline MTCQ1^mir21–^/–. Right, quantitative real-time PCR for analyzing the expression of mir-21 of MTCQ-WT and MTCQ1^mir21–^/–. Data represent means±SD. ***p<0.001 by Student’s t-test. n=3 independent experiments (each experiment contains two technical replicates). (B) Schema of the experiment for assaying inflammasome activities of murine tumors. 1×10⁶ of MTCQ1-WT or MTCQ1^mir21–^/– cells were inoculated to the subcutaneous region of the wild-type C57BL/6J mice for 2 weeks. Cisplatin 5 mg/kg was given intraperitoneally for 4 consecutive days. The mice were sacrificed on the fifth day after the start of cisplatin injection. F4/80⁺ tumor associated macrophages (TAMs) were harvested and caspase 1 activity (FLICA⁺) was analyzed by flow cytometry. (C) Left, representative data of the flow cytometry analysis for the caspase 1 activity of the TAMs (F4/80⁺FLICA⁺) after treated with cisplatin or the control PBS. n=5 for each group. Data shows mean±SD. *p<0.05 by Student’s t-test. (D) Schema of the experiment of (E) and (F). 1×10⁶ of wild-type MTCQ1 cells (MTCQ1-WT) or MTCQ1^mir21−^/− cells were inoculated to the subcutaneous region of the wild-type or Nlrp3^−/− C57BL/6J mice. After the tumor size reached 50 mm³ (day 0), intraperitoneal injection of cisplatin (50 mg/kg) or PBS was given every 3 days for a total of six doses, and the tumor size were measured every 3 days. Mice were sacrificed at the 18th day and tumor weight were measured. (E) The volume the MTCQ1-WT/MTCQ1^mir21−/−-formed tumors in wild-type or Nlrp3⁻^/− mice treated with cisplatin or PBS. n=5 for each group. Data shows mean±SD. **p<0.01, ns=no significance by Student’s t-test. (F) The waterfall plots to indicate the tumor volume change of each mouse. (G) The weight of the MTCQ1-WT/MTCQ1^mir21−/−-formed tumors in wild-type or Nlrp3^−/− mice treated with cisplatin or PBS. n=5 for each group. Data shows mean±SD. **p<0.01 ns=no significance by Student’s t-test. miR, micro RNA; PBS, phosphate buffered saline.