Journal of Traditional Chinese Medicine ›› 2022, Vol. 42 ›› Issue (5): 693-700.DOI: 10.19852/j.cnki.jtcm.2022.05.003
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Qilan preparation (芪蓝颗粒) inhibits proliferation and induces apoptosis by down-regulating microRNA-21 in human Tca8113 tongue squamous cell carcinoma cells
DING Jiamin1, XING Yifeng2, CHEN Zuoliang3, CHEN Wanlu1, MA Zhongxiong1, XIE Yunde1, ZHOU Lin4(
)
- 1 Department of Oral Mucosal Diseases, Affiliated Stomatological Hospital of Fujian Medical University, Fuzhou 350000, China
2 School and Hospital of Stomatology, Fujian Medical University, Fuzhou 350000, China
3 Department of Oral Mucosal Diseases, Xiamen Stomatological Hospital, School of Stomatology, Fujian Medical University, Xiamen 361003, China
4 Department of Implantology, Affiliated Stomatological Hospital of Fujian Medical University, Fuzhou 350001, China
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Received:2021-05-15Accepted:2021-08-08Online:2022-09-02Published: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: +86-591-83754882
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Supported by:Fujian Provincial Healthy and Family Planning Commission Program: Effects of Qilan Preparation on the Expression of miRNA-21 and Its Target Proteins in Human Tongue Squamous Cell Carcinoma Cells(2017-2-38)
Cite this article
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.
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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 |
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 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 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.
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.
| 1. | Chen ZL, Chen HB. Clinical study on the effect of 1023 traditional Chinese medicine mixture on the treatment of oral precancerous lesions and cancer prevention. Lin Chuan Kou Qiang Yi Xue Za Zhi 2002; 18: 458-9. |
| 2. |
Hou J, Liu S, Ma Z, et al. Effects of gynostemma pentaphyllum makino on the immunological function of cancer patients. J Tradit Chin Med 1991; 11: 47-52.
PMID |
| 3. |
Chen JC, Lu KW, Tsai ML, et al. Gypenosides induced G0/G1 arrest<i>via</i> CHk2 and apoptosis through endoplasmic reticulum stress and mitochondria-dependent pathways in human tongue cancer SCC-4 cells. Oral Oncol 2009; 45: 273-83.
DOI URL |
| 4. | Chen ZL, Guan YQ, Li HF. The orthogonal design study on tumor inhibition and cancer block role of Chinese medicinal 1023 co. Zhong Hua Kou Qiang Yi Xue Za Zhi 1998; 33: 50-2. |
| 5. |
Medina PP, Nolde M, Slack FJ. OncomiR addiction in an in vivo model of microRNA-21-induced pre-B-cell lymphoma. Nature 2010; 467: 86-90.
DOI URL |
| 6. |
Li J, Huang H, Sun L, et al. MiR-21 indicates poor prognosis in tongue squamous cell carcinomas as an apoptosis inhibitor. Clin Cancer Res 2009; 15: 3998-4008.
DOI URL |
| 7. |
Ren W, Wang X, Gao L, et al. MiR-21 modulates chemosensitivity of tongue squamous cell carcinoma cells to cisplatin by targeting PDCD4. Mol Cell Biochem 2014; 390: 253-62.
DOI URL |
| 8. |
Zhou X, Ren Y, Liu A, et al. STAT3 inhibitor WP1066 attenuates miRNA-21 to suppress human oral squamous cell carcinoma growth in vitro and in vivo. Oncol Rep 2014; 31: 2173-80.
DOI URL |
| 9. |
Mestdagh P, Van VP, De Weer A, et al. A novel and universal method for microRNA RT-qPCR data normalization. Genome Biol 2009; 10: R64.
DOI URL |
| 10. |
Brito JA, Gomes CC, Guimarães AL, Campos K, Gomez RS. Relationship between microRNA expression levels and histopathological features of dysplasia in oral leukoplakia. J Oral Pathol Med 2014; 43: 211-6.
DOI PMID |
| 11. | Mo Q, Zhou G, Xie B, et al. Evaluation of the hepatoprotective effect of Yigan mingmu oral liquid against acute alcohol-induced liver injury in rats. BMC Complement Med 2020; 20: 32. |
| 12. |
Chen J, Wu A, Bamodu O, et al. Ovatodiolide suppresses oral cancer malignancy by down-regulating exosomal Mir-21/STAT3/β-catenin cargo and preventing oncogenic trans-formation of normal gingival fibroblasts. Cancers 2019; 12: 56
DOI URL |
| 13. |
Lim LP, Lau NC, Garrett-Engele P, et al. Microarray analysis shows that some microRNAs downregulate large numbers of target mRNAs. Nature 2005; 433: 769-73.
DOI URL |
| 14. |
Shibahara K, Asano M, Ishida Y, et al. Isolation of a novel mouse gene MA-3 that is induced upon programmed cell death. Gene 1995; 166: 297-301.
PMID |
| 15. |
Ye Y, Yang S, Han Y, et al. HOXD-AS1 confers cisplatin resistance in gastric cancer through epigenetically silencing PDCD4 via recruiting EZH2. Open biol 2019; 9: 190068
DOI URL |
| 16. |
Gao F, Wang X, Zhu F, et al. PDCD4 gene silencing in gliomas is associated with 5′ CpG island methylation and unfavourable prognosis. J Cell Mol Med 2009; 13: 4257-67.
DOI URL |
| 17. |
Parsons R. Human cancer, PTEN and the PI-3 kinase pathway. Semin Cell Dev Biol 2004; 15: 171-6.
PMID |
| 18. |
Sansal I, Sellers WR. The biology and clinical relevance of the PTEN tumor suppressor pathway. J Clin Oncol 2004; 22: 2954-63.
DOI URL |
| 19. |
Wang J, Li Y, Wang X, Jiang C. Ursolic acid inhibits proliferation and induces apoptosis in human glioblastoma cell lines U251 by suppressing TGF-β1/miR-21/PDCD4 pathway. Basic Clin Pharmacol 2012; 111: 106-12.
DOI PMID |
| 20. |
Hu HY, Li KP, Wang XL, et al. Set9, NF-κB, and microRNA-21 mediate berberine-induced apoptosis of human multiple myeloma cells. Acta Pharmacol Sin 2012; 34: 157-66.
DOI URL |
| 21. |
Li LQ, Li XL, Wang L, et al. Matrine inhibits breast cancer growth via miR-21/PTEN/Akt pathway in MCF-7 cells. Cell Physiol Biochem 2012;30: 631-41.
DOI URL |
| 22. | Ma W, Hu S, Xu Y, et al. Haimufang decoction, a Chinese medicine formula for lung cancer, arrests cell cycle, stimulates apoptosis in NCI-H1975 cells, and induces M1 polarization in RAW 264.7 macrophage cells. BMC Complement Med 2020; 20: 243. |
| 23. |
Wei ZT, Zhang X, Wang XY, et al. PDCD4 inhibits the malignant phenotype of ovarian cancer cells. Cancer Sci 2009; 100: 1408-13.
DOI URL |
| 24. |
Lankat-Buttgereit B, Lenschen B, Schmidt H, Göke R. The action of Pdcd4 may be cell type specific: evidence that reduction of dUTPase levels might contribute to its tumor suppressor activity in Bon-1 cells. Apoptosis 2008; 13: 157-64.
PMID |
| 25. |
Jansen AP, Camalier CE, Stark C, Colburn NH. Characterization of programmed cell death 4 in multiple human cancers reveals a novel enhancer of drug sensitivity. Mol Cancer Ther 2004; 3: 103-10.
PMID |
| 26. |
Yang HS, Matthews CP, Clair T, et al. Tumorigenesis suppressor Pdcd4 down-regulates mitogen-activated protein kinase kinase kinase kinase 1 expression to suppress colon carcinoma cell invasion. Mol Cell Biol 2006; 26: 1297-306.
DOI URL |
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