敲降miR-222-3p靶向PTEN对甲状腺癌131I放疗抵抗的影响及其机制

doi:10.13481/j.1671-587X.20210318

Abstract

Abstract: Objective

To investigate the effect of miR-222-3p on the proliferation and apoptosis of thyroid cancer cells via regulating gene of phosphatase and tension homolog deleted on chromosome ten（PTEN），and to elucidate the role of miR-222-3p in ¹³¹I radiotherapy resistance of thyroid cancer cells and its mechanism.

Methods

The expression levels of miR-222-3p in the human thyroid cancer cells and human normal thyroid follicular epithelial Nth-ori 3-1 cells were determined by Real-time fluorescence quantitative PCR（RT-qPCR） method.The thyroid cancer K1 cells and ¹³¹I resistance of thyroid cancer K1R cells were used as the subjects and were divided into blank control group， ¹³¹I treatment group， ¹³¹I+miR-222-3p knockdown group， ¹³¹I+PTEN over-expression group，and ¹³¹I+PTEN over-expression+miR-222-3p over-expression group. The K1 cells and KIR cells in blank control group didn’t receive any treatment； both the K1 and K1R cells in ¹³¹I treatment group were treated with ¹³¹I at doses of 1 and 3 Gy； the cells in ¹³¹I+miR-222-3p knockdown group were transfected with miR-222-3p inhibitor prior to ¹³¹I treatment at dose of 1 and 3 Gy； the cells in ¹³¹I+PTEN over-expression group were transfected with pcDNA3.1-PTEN prior to ¹³¹I treatment with doses of 1 and 3 Gy； the cells in ¹³¹I+PTEN over-expression+miR-222-3p over-expression group were simultaneously transfected with pcDNA3.1-PTEN and miR-222-3p mimic prior to ¹³¹I treatment with doses of 1 and 3 Gy. Clony formation assay was used to detect the clony formation number of K1 and K1R cells；CCK-8 assay was used to detect the proliferation activities of K1 and K1R cells；Annexin Ⅴ-FITC / PI double staining method was used to detect the apoptotic rates of K1 and K1R cells；Western blotting method was used to detect the expression levels of PTEN protein in K1 and K1R cells； Double luciferase reporter gene assay was used to verify the targeting relationship between miR-222-3p and PTEN.

Results

Compared with the Nth-ori 3-1 cells，the expression levels of miR-222-3p in thyroid carcinoma cells were increased （P<0.01）； the expression level of miR-222-3p in the thyroid cancer K1R cells was higher than that in thyroid cancer K1 cells （P<0.01）. Compared with ¹³¹I treatment group， the clony formation number of the K1 and K1R cells in ¹³¹I + miR-222-3p knockdown group was decreased（P<0.01）， the proliferation activities of K1 and K1R cells were decreased （P<0.01），and the apopotic rates were increased（P<0.01）. Compared with miR-NC-PTEN-WT group， the luciferase activity of HEK-293 cells in miR-222-3P-PTEN-WT group was decreased （P<0.01）. Compared with before knockdown， the expression level of PTEN protein in K1 and K1R cells were significantly increased after knockdown（P<0.01）. Compared with ¹³¹I + PTEN over-expression group， the colony formation number of K1 and K1R cells in ¹³¹I +PTEN over-expression+miR-222-3p over-expression group was increased （P<0.01）， the proliferation activities were increased （P<0.01）， and the apoptotic rate was decreased （P<0.01）.

Conclusion

Knockdown of miR-222-3p targeting PTEN and up-regulating its expression level can alleviate the ¹³¹I radiotherapy resistance of thyroid cancer cells.

Key words: miR-222-3p, gene of phosphatase and tension homolog deleted on chromosome ten, thyroid neoplasms, ¹³¹I radiotherapy resistance

CLC Number:

R736.1

Yaoyu FENG, Chenglei ZHANG, Lijuan HOU, Yifu WANG, Xiuling WU, Yunhai MA. Effect of miR-222-3p knockdown targeting PTEN on ¹³¹I radiotherapy resistance of thyroid cancer and its mechanism[J].Journal of Jilin University(Medicine Edition), 2021, 47(3): 677-686.

Figures/Tables 15

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Primer	Sequence
miR-222-3p	F: 5'-GGGGAGCTACATCTGGCT-3'
miR-222-3p	R: 5'-TGCGTGTCGTGGAGTC-3'
U6	F: 5'-GCTTCGGCAGCACATATACTAAAAT-3'
U6	R: 5'-CGCTTCACGAATTTGCGTGTCAT-3'

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Effect of miR-222-3p knockdown targeting PTEN on ¹³¹I radiotherapy resistance of thyroid cancer and its mechanism

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Effect of miR-222-3p knockdown targeting PTEN on 131I radiotherapy resistance of thyroid cancer and its mechanism

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Effect of miR-222-3p knockdown targeting PTEN on ¹³¹I radiotherapy resistance of thyroid cancer and its mechanism