Journal of Traditional Chinese Medicine ›› 2022, Vol. 42 ›› Issue (5): 693-700.DOI: 10.19852/j.cnki.jtcm.2022.05.003
• Research Articles • Previous Articles Next Articles
DING Jiamin1, XING Yifeng2, CHEN Zuoliang3, CHEN Wanlu1, MA Zhongxiong1, XIE Yunde1, ZHOU Lin4()
Received:
2021-05-15
Accepted:
2021-08-08
Online:
2022-09-02
Published:
2022-09-02
Contact:
ZHOU Lin
About author:
ZHOU Lin, Department of Implantology, Affiliated Stomatological Hospital of Fujian Medical University, Fuzhou 350001, China. zhoulin@fjmu.edu.cnTelephone: +86-591-83754882Supported by:
DING Jiamin, XING Yifeng, CHEN Zuoliang, CHEN Wanlu, MA Zhongxiong, XIE Yunde, ZHOU Lin. Qilan preparation (芪蓝颗粒) inhibits proliferation and induces apoptosis by down-regulating microRNA-21 in human Tca8113 tongue squamous cell carcinoma cells[J]. Journal of Traditional Chinese Medicine, 2022, 42(5): 693-700.
Concentrations (mg/mL) | 12 h | 24 h | 48 h |
---|---|---|---|
Control | 100.00±1.42 | 100.00±1.29 | 100.00±4.82 |
1.56 | 92.66±2.60a | 81.41±1.19b | 66.22±2.65c |
3.12 | 78.70±2.16a | 68.20±1.55b | 57.24±1.90c |
6.25 | 65.31±2.00a | 54.59±2.45b | 43.79±1.22c |
12.5 | 40.91±0.83a | 31.68±2.76b | 12.22±0.48c |
25 | 10.54±0.98a | 6.03±1.12b | 4.19±0.15c |
SUM | 64.69±31.82 | 56.98±32.07 | 47.28±33.50 |
F value | 1080.527 | 1028.074 | 640.405 |
P value | 0.000 | 0.000 | 0.000 |
Table 1 Effect of Qilan preparation on Tca8113 cell viability in vitro
Concentrations (mg/mL) | 12 h | 24 h | 48 h |
---|---|---|---|
Control | 100.00±1.42 | 100.00±1.29 | 100.00±4.82 |
1.56 | 92.66±2.60a | 81.41±1.19b | 66.22±2.65c |
3.12 | 78.70±2.16a | 68.20±1.55b | 57.24±1.90c |
6.25 | 65.31±2.00a | 54.59±2.45b | 43.79±1.22c |
12.5 | 40.91±0.83a | 31.68±2.76b | 12.22±0.48c |
25 | 10.54±0.98a | 6.03±1.12b | 4.19±0.15c |
SUM | 64.69±31.82 | 56.98±32.07 | 47.28±33.50 |
F value | 1080.527 | 1028.074 | 640.405 |
P value | 0.000 | 0.000 | 0.000 |
Figure 1 Effect of Qilan preparation on Tca8113 cell apoptosis and cell cycle Tca8113 cells were treated without (control) and with Qilan preparation (1.56, 3.12, 6.25, 12.5, and 25 mg/mL) for 24 h. The cells were stained with fluorescein isothiocyanate (FITC) -conjugated Annexin V and propidium iodide (PI) for flow cytometry. A: the cell populations shown in the upper (Annexin V+/PI+) and lower (Annexin V+/PI-) right represent the apoptotic cells. B: the percentage of apoptotic cells, aP < 0.05 vs Control. Tca8113 cells were treated without (Control) and with Qilan preparation (1.56, 3.12, 6.25, 12.5, and 25 mg/mL) for 12 h. Next, the cells were stained with PI for flow cytometry. C, D: the cell cycle distribution (G0/G1, S, and G2/M) was based on 2N and 4N deoxyribonucleic acid (DNA) content for the DNA content analysis using the FCS Express 4 plus software. The data are represented as the mean ± standard deviation for three experiments per group. aP < 0.05 vs Control for S-phase percentage; bP < 0.05 vs Control for G1/G0-phase percentage.
Figure 2 Effect of Qilan preparation on mir-21 expression and PDCD4 and PTEN in Tca8113 cells Expression of miR-21, programmed cell death 4 (PDCD4) and phosphatase and gensin homologue (PTEN) were separately detected by stem-loop a Taqman assay and Western blot following treatment without (Control) and with Qilan preparation (1.56, 3.12, 6.25, 12.5, and 25 mg/mL) for 24 h. A: the fold change in relative expression of miR-21 to small nucleolar RNA, C/D box 44 (RNU44). The fold change in relative expression of (B) PDCD4 or (C) PTEN to glyceraldehyde-3-phosphate dehydrogenase (GAPDH). The data are represented as the mean ± standard deviation for three experiments per group. aP < 0.05 vs Control.
Figure 3 Downregulation of miR-21 partially contributes to Qilan preparation-induced cell growth inhibition and apoptosis induction The miR-21 mimic and negative control ribonucleic acid (NC RNA, which had the same number of basic groups but a different sequence compared with the miR-21 mimic) were introduced into the Tca8113 cells for 24 h. Transfected or non-transfected cells were treated with 6.25 mg/mL of the Qilan preparation for 24 h. Thus, there were five groups including Control, NC RNA, Qilan preparation, NC RNA + Qilan preparation and miR-21 + Qilan preparation group. Basic medium was used as a blank control (Control group). NC RNA group was used as a negative control. Qilan preparation group was used as a positive control. Significant differences were observed in the miR-21 expression levels between the miR-21 + Qilan preparation, the NC RNA and the NC RNA + Qilan preparation groups (A). After the level of miR-21 was increased, (B) the cell viability increased, but the (C and D) apoptosis ratio and (E) programmed cell death 4 (PDCD4) expression levels decreased. The data are represented as the mean ± standard deviation for three or four experiments. There were no significant differences between Control and NC RNA groups. aP < 0.05 vs the NC RNA group and bP < 0.05 vs the NC RNA + Qilan preparation group.
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