Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2020 Jun;43(6):1906-1914.
doi: 10.3892/or.2020.7547. Epub 2020 Mar 17.

miR‑10a increases the cisplatin resistance of lung adenocarcinoma circulating tumor cells via targeting PIK3CA in the PI3K/Akt pathway

Tonghai Huang  1 , Kangqi Ren  1 , Guanggui Ding  1 , Lin Yang  1 , Yuxin Wen  1 , Bin Peng  1 , Guangsuo Wang  1 , Zheng Wang  1
Affiliations

miR‑10a increases the cisplatin resistance of lung adenocarcinoma circulating tumor cells via targeting PIK3CA in the PI3K/Akt pathway

Tonghai Huang et al. Oncol Rep. 2020 Jun.

Abstract

Circulating tumor cells (CTCs) that are shed from the primary tumor invade the blood stream or surrounding parenchyma to form new tumors. The present study aimed to explore the underlying mechanism of cisplatin resistance in lung adenocarcinoma CTCs and provide clinical treatment guidance for lung cancer treatment. CTCs from the blood samples of 6 lung adenocarcinoma patients were treated with different concentrations of cisplatin along with A549 and H1299 cells. The sensitivity of CTCs to cisplatin was explored by detecting the inhibitory rate via CCK‑8 assay. The related molecular mechanism was investigated by western blot analysis. miR‑10a expression was detected using quantitative real‑time PCR (RT‑qPCR). The relationship between miR‑10a and phosphatidylinositol‑4,5‑bisphosphate 3‑kinase catalytic subunit α (PIK3CA) was verified and further confirmed by luciferase reporter assay, western blotting and RT‑qPCR assay. The results revealed that CTCs exhibited lower cisplatin sensitivity than A549 and H1299 cells. Moreover, CTCs treated with cisplatin demonstrated higher miR‑10a expression and lower PIK3CA expression than that in A549 and H1299 cells (P<0.01). Expression of phosphoinositide 3‑kinase (PI3K) and protein kinase B (Akt) phosphorylation were also decreased in A549 and H1299 cells compared with CTCs after cisplatin treatment. PIK3CA is a target of miR‑10a, and both miR‑10a overexpression and PIK3CA knockdown obviously decreased the sensitivity of A549 and H1299 cells to cisplatin as well as the expression of PI3K and phosphorylation of Akt. PIK3CA overexpression attenuated the cisplatin resistance of A549 and H1299 cells induced by miR‑10a. In conclusion, miR‑10a suppressed the PI3K/Akt pathway to strengthen the resistance of CTCs to cisplatin via targeting PIK3CA, providing a new therapeutic target for lung cancer treatment.

Keywords: lung adenocarcinoma; circulating tumor cells; miR-10a; PIK3CA; cisplatin resistance.

PubMed Disclaimer

Figures

Figure 1.
Figure 1.
Inhibitory rates for cisplatin treatment in lung adenocarcinoma A549 and H1299 cells and CTCs from 6 patients. CTCs, circulating tumor cells; IC50, half maximal inhibitory concentration.
Figure 2.
Figure 2.
Expression of miR-10a and PIK3CA in lung adenocarcinoma A549 and H1299 cells and CTCs from 6 patients. (A) Expression of miR-10a was determined by RT-qPCR in A549 and A1299 cells and CTCs. (B) Expression of PIK3CA and the activation of the PI3K/Akt signaling pathway were determined by western blotting in A549 and A1299 cells and CTCs. CTCs were isolated from the blood samples obtained from 6 lung adenocarcinoma patients. **P<0.01 and ***P<0.001 compared with A549 cells; ##P <0.01 and ###P<0.001 compared with H1299 cells. PIK3CA, phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit α; PI3K, phosphoinositide 3-kinase; pAkt, phosphorylated protein kinase B; CTCs, circulating tumor cells.
Figure 3.
Figure 3.
PIK3CA is a target of miR-10a. (A) Prediction of the target regulatory relationship between PIK3CA and miR-10a. (B) Regulatory relationship was confirmed by luciferase reporter assay. WT, wild-type, MUT, mutant; NC, negative control; miR-10a, miR-10a mimic. **P<0.01 compared with the NC group. (C and D) A549 cells were transfected with the miR-10a inhibitor, and expression of PIK3CA was detected by western blotting and real-time PCR, respectively. GAPDH was used as an internal control. ***P<0.001 compared with the NC group. PIK3CA, phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit α.
Figure 4.
Figure 4.
Overexpression of miR-10a enhances the cisplatin resistance of lung adenocarcinoma A549 and H1299 cells. (A) Cisplatin-mediated inhibitory rate of A549 cells after miR-10a overexpression or PIK3CA knockdown (shPIK3CA). (B) Cisplatin-mediated inhibitory rate of H1299 after miR-10a overexpression or PIK3CA knockdown. Expression of PIK3CA and the activation of the PI3K/Akt signaling pathway were determined by western blotting in (C and D) A549 and (E and F) H1299 cells after transfection with miR-10a mimics or shPIK3CA. **P<0.01 compared with the NC group. PIK3CA, phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit α; PI3K, phosphoinositide 3-kinase; pAkt, phosphorylated protein kinase B.
Figure 5.
Figure 5.
Overexpression of PIK3CA reduces the effect of miR-10a on cisplatin resistance. (A) Expression of PIK3CA in A549 and H1299 cells after overexpression of miR-10a and PIK3CA. (B) Quantitative expression of PIK3CA in A549 and H1299 cells after overexpression of miR-10a and PIK3CA. Cisplatin-mediated inhibitory rate of (C) A549 and (D) H1299 cells after overexpressing miR-10a and PIK3CA. **P<0.01 compared with the NC group. PIK3CA, phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit α.
See this image and copyright information in PMC

References

    1. Siegel RL, Miller KD, Jemal A. Cancer Statistics, 2017. CA Cancer J Clin. 2017;67:7–30. doi: 10.3322/caac.21387. - DOI - PubMed
    1. Sorber L, Zwaenepoel K, Deschoolmeester V, Van Schil PE, Van Meerbeeck J, Lardon F, Rolfo C, Pauwels P. Circulating cell-free nucleic acids and platelets as a liquid biopsy in the provision of personalized therapy for lung cancer patients. Lung Cancer. 2017;107:100–107. doi: 10.1016/j.lungcan.2016.04.026. - DOI - PubMed
    1. Gan Y, Zhou P, Li W. D62 types, Genotypes, and Phenotypes: The Three Ts of thoracic oncology. American Thoracic Society; 2018. Correlations of Tumor Stage and Serum Tumor Markers with Age in Nonsmoking Females with Lung Adenocarcinoma; p. ppA7334.
    1. Zhai X, Zheng Q, Yang L, Zhu Y, Li J, Liu Y, Wang Y. Impact of platinum/pemetrexed combination versus other platinum-based regimens on adjuvant chemotherapy in resected lung adenocarcinoma. Sci Rep. 2017;7:1453. doi: 10.1038/s41598-017-01347-6. - DOI - PMC - PubMed
    1. Jin B, Niu Y, Zhang Y, Chu T, Gu A, Wu J, Pei J, Zhu L, Han B. 1281Pcombination of chemotherapy and gefitinib as first-line treatment of patients with advanced lung adenocarcinoma and sensitive egfr mutations: A randomised controlled trial. Ann Oncol. 2014;25(4 Suppl):iv451. doi: 10.1093/annonc/mdu349.62. - DOI - PubMed

MeSH terms