南方医科大学学报

南方医科大学学报 ›› 2020, Vol. 40 ›› Issue (05): 698-702.doi: 10.12122/j.issn.1673-4254.2020.05.13

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长链非编码RNA MALAT1靶向miR-485-3p调控乳腺癌细胞对紫杉醇的耐药性

艾尼·沙塔尔，闫焕英，丁 伟，阿迪力江，苏鹏程   

  • 出版日期:2020-05-20 发布日期:2020-05-20
  • 基金资助:

Long-chain non-coding RNA MALAT1 regulates paclitaxel resistance of breast cancer cells by targeting miR-485-3p

  

  • Online:2020-05-20 Published:2020-05-20

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摘要： 目的 研究长链非编码RNAMALAT1对乳腺癌细胞紫杉醇耐药性的影响机制。方法 运用紫杉醇浓度梯度诱导法检测紫杉醇耐药乳腺癌细胞SK-BR-3；将si-NC组（转染si-NC）、si-MALAT1组（转染si-MALAT1）、pc DNA组（转染pc DNA）、pcDNAMALAT1组（转染pc DNA-MALAT1）、miR-NC组（转染miR-NC）、miR-485-3p组（转染miR-485-3p mimics）、si-MALAT1+antimiR-NC组（共转染si-MALAT1和anti-miR-NC）、si-MALAT1+anti-miR-485-3p组（共转染si-MALAT1和anti-miR-485-3p），均用脂质体法转染SK-BR-3/PR细胞；MTT法检测细胞IC50；Western blot检测细胞中P-gp、Bax、Bcl-2的蛋白表达；双荧光素酶报告基因检测实验检测细胞中MALAT1与miR-485-3p的结合力。结果 与紫杉醇敏感SK-BR-3细胞相比， SK-BR-3/PR细胞的IC50、MALAT1均上调，miR-485-3p下调，差异具有统计学意义（P<0.05）。沉默MALAT1或过表达miR-485-3p均可下调SK-BR-3/PR细胞的IC50，下调P-gp和Bcl-2表达，上调Bax表达，差异具有统计学意义（P<0.05）。miR-485-3p是MALAT1的靶标。抑制miR-485-3p可逆转沉默MALAT1对SK-BR-3/PR细胞的IC50、P-gp、Bcl-2和Bax表达的影响，差异具有统计学意义（P<0.05）。结论 长 链非编码RNAMALAT1可调控乳腺癌细胞对紫杉醇的耐药性，其机制可能与靶向miR-485-3p下调P-gp、Bcl-2，上调Bax有关。

Abstract: Objective To investigate the role of long-chain non-coding RNA MALAT1 in modulating paclitaxel resistance in breast cancer cells. Methods Breast cancer SK-BR-3 cells were treated with gradient concentrations of paclitaxel to induce paclitaxel resistance of the cells. The resistant cells were transfected with si-NC, si-MALAT1, pcDNA, pcDNA-MALAT1, miRNC, miR-485-3p mimics, si-MALAT1+anti-miR-NC, or si-MALAT1+anti-miR-485-3p via liposomes. Following the transfections, the cells were examined for changes in IC50 of paclitaxel using MTT assay; the protein expression of P-gp, Bcl-2 and Bax were detected with Western blotting, and a dual luciferase reporter assay was used to detect the binding of MALAT1 to miR-485-3p. Results Compared with paclitaxel-sensitive SK-BR-3 cells, paclitaxel-resistant SK-BR-3 cells showed significantly increased the IC50 of paclitaxel with up-regulated MALAT1 expression and down-regulated miR- 485- 3p expression (P<0.05). Silencing MALAT1 or overexpressing miR- 485-3p obviously lowered the IC50 of paclitaxel and the expression of P-gp and Bcl-2 and increased the expression of Bax in SK-BR-3/PR cells (P<0.05). miR-485-3p was identified as the target of MALAT1, and inhibiting miR-485-3p significantly reverse the effect of MALAT1 silencing on IC50 of paclitaxel and the expressions of P-gp, Bcl- 2 and Bax in SK-BR-3/PR cells (P<0.05). Conclusion MALAT1 can modulate paclitaxel resistance in breast cancer cells possibly by targeting miR-485-3p to down-regulate P-gp and Bcl-2 and up-regulate Bax.