J Breast Cancer.  2018 Sep;21(3):251-258. 10.4048/jbc.2018.21.e40.

Hsa-miRNA-143-3p Reverses Multidrug Resistance of Triple-Negative Breast Cancer by Inhibiting the Expression of Its Target Protein Cytokine-Induced Apoptosis Inhibitor 1 In Vivo

• Deng YW
• Hao WJ
• Li YW
• Li YX
• Zhao BC
• Lu D

• KMID: 2421365
• DOI: http://doi.org/10.4048/jbc.2018.21.e40

Abstract

PURPOSE
Multidrug resistance (MDR) remains a major obstacle in the treatment of triple-negative breast cancer (TNBC) with conventional chemotherapeutic agents. A previous study demonstrated that hsa-miRNA-143-3p plays a vital role in drug resistance of TNBC. Downregulation of hsa-miRNA-143-3p upregulated the expression of its target protein cytokine-induced apoptosis inhibitor 1 (CIAPIN1) in order to activate MDR, while upregulation of hsa-miRNA-143-3p effectively enhances the sensitivity of drug-resistant TNBC cells to chemotherapeutics. The present study aimed to further verify these findings in vivo.
METHODS
We established a hypodermic tumor nude mice model using paclitaxel-resistant TNBC cells. We expressed ectopic hsa-miRNA-143-3p under the control of a breast cancer-specific human mammaglobin promoter that guided the efficient expression of exogenous hsa-miRNA-143-3p only in breast cancer cells. Thereafter, we overexpressed hsa-miRNA-143-3p in xenografts using a recombinant virus system and quantified the expression of hsa-miRNA-143-3p, CIAPIN1 protein, and proteins encoded by related functional genes by western blot.
RESULTS
We successfully completed the prospective exploration of the intravenous virus injection pattern from extensive expression to targeted expression. The overexpression of hsa-miRNA-143-3p significantly alleviated chemoresistance of TNBC by inhibiting viability. In addition, we observed that the expression of CIAPIN1 as a hsa-miRNA-143-3p target protein was remarkably decreased.
CONCLUSION
We partly illustrated the mechanism underlying the hsa-miRNA-143-3p/CIAPIN1 drug resistance pathway. HsamiRNA-143-3p as a tumor suppressive microRNA may be a novel target to effectively reverse MDR of TNBC in vivo.

Keyword

Cytokine-induced apoptosis inhibitor 1; Human mammaglobin; Micro-ribonucleic acid-143-3p; Multiple drug resistance; Triple negative breast neoplasms

MeSH Terms

Animals
Apoptosis*
Blotting, Western
Breast
Breast Neoplasms
Down-Regulation
Drug Resistance
Drug Resistance, Multiple*
Heterografts
Humans
Mice
Mice, Nude
MicroRNAs
Prospective Studies
Triple Negative Breast Neoplasms*
Up-Regulation
MicroRNAs

Figure

• 

  Figure 1 Detection of expression efficiency and specificity of hsa-miRNA-143-3p under control of the hMAM promoter. (A) The recombinant viruses were Lv-NC, Lv-miRNA-143-3p (containing the CMV promoter), and Lv-hMAM-miRNA-143-3p (containing the hMAM promoter). For 72 hours after infection of MDA-MB-231 cells, the relative content of hsa-miRNA-143-3p is shown, with U6 level as a reference. Relative hsa-miRNA-143-3p values were normalized to that of the control group as a reference for data comparison. (B) MDA-MB-231, BT-20, MCF7, SKBr-3, Lovo, HepG2, and A549 cells were infected with Lv-NC, Lv-miRNA-143-3p, or Lv-hMAM-miRNA-143-3p, respectively. After 72 hours, the relative content of hsa-miRNA-143-3p was determined, with U6 level as a reference. Relative hsa-miRNA-143-3p values were normalized to that of the control group as a reference for data comparison. The experimental design was repeated 3 times. Data are represented as mean±standard error of the mean.Lv-hMAM-miRNA-143-3p=lentiviral-expressed vector human mammaglobin (hMAM) miRNA-143-3p; Lv-NC=lentiviral-expressed vector no-antisense oligomers control group; CMV=cytomegalovirus. *p=0.001 vs. control group.

• 

  Figure 2 Overexpression of hsa-miRNA-143-3p and the sensitivity of triple-negative breast cancer to paclitaxel. (A) The tumor volume growth rate and tumor size in the Lv-hMAM-miRNA-143-3p+paclitaxel group were significantly reduced compared with those in the Lv-NC+paclitaxel group, paclitaxel group, and physiological saline control group. (B) Upregulating the expression of hsa-miRNA-143-3p prominently increased the tumor growth inhibition ratio in the Lv-hMAM-miRNA-143-3p+paclitaxel group compared to that of the other groups. The experimental design was repeated 3 times. Data are represented as mean±standard error of the mean.Lv-hMAM-miRNA-143-3p=lentiviral-expressed vector human mammaglobin (hMAM) miRNA-143-3p; Lv-NC=lentiviral-expressed vector no-antisense oligomers control group. *p=0.001.

• 

  Figure 3 Activation of multidrug resistance by cytokine-induced apoptosis inhibitor 1 (CIAPIN1) targeting to hsa-miRNA-143-3p in triple-negative breast cancer (TNBC). (A) Transcript level of hsa-miRNA-143-3p notably increased upon infection with hsa-miRNA-143-3p compared with the level in other groups as determined by real-time polymerase chain reaction. (B) Western blot showing that CIAPIN1 protein levels decreased in TNBC tissue expressing hsa-miRNA-143-3p. The experimental design was repeated 3 times. Data are represented as mean±standard error of the mean.β-actin=reference protein; Lv-hMAM-miRNA-143-3p=lentiviral-expressed vector hMAM miRNA-143-3p; Lv-NC=lentiviral-expressed vector no-antisense oligomers control group. *p=0.001.

• 

  Figure 4 Hsa-miRNA-143-3p expression and other drug resistance-relating proteins. Western blot showing that P53 protein levels increased while multidrug resistance gene 1 (MDR1) levels decreased in the Lv-hMAM-miRNA-143-3p+paclitaxel group compared to levels in other groups. The experimental design was repeated 3 times. Data are represented as mean±standard error of the mean. (A) Western blot showing that P53 protein levels increased while MDR1 levels decreased in the Lv-hMAM-miRNA-143-3p+paclitaxel group compared to levels in other groups. (B) Electrophoretic band of the P53 and MDR1 expression level as described above.β-actin=reference protein; Lv-hMAM-miRNA-143-3p=lentiviral-expressed vector hMAM miRNA-143-3p; Lv-NC=lentiviral-expressed vector no-antisense oligomers control group. *p=0.001.

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