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Article Navigation >  Journal of Pharmaceutical Practice and Service  > 2019  >  37(5): 433-439

ZHOU Jing, WU Gao, MA Fujia. Study on the mechanism of proliferation of human lung cancer A549 cells regulated by hsa-miRNA-30a-5p[J]. Journal of Pharmaceutical Practice and Service, 2019, 37(5): 433-439. doi: 10.3969/j.issn.1006-0111.2019.05.009
Citation: ZHOU Jing, WU Gao, MA Fujia. Study on the mechanism of proliferation of human lung cancer A549 cells regulated by hsa-miRNA-30a-5p[J]. Journal of Pharmaceutical Practice and Service, 2019, 37(5): 433-439. doi: 10.3969/j.issn.1006-0111.2019.05.009
shu

Study on the mechanism of proliferation of human lung cancer A549 cells regulated by hsa-miRNA-30a-5p

doi: 10.3969/j.issn.1006-0111.2019.05.009
  • 1.

    Department of Pharmacy, Ningbo Women's and Children's Hospital, Ningbo, 315012, China

  • 2.

    Hongkou Branch, Changhai Hospital Affiliated to Naval Medical University, Shanghai, 200081, China

  • 3.

    Department of Pharmacy, Naval Special Medical Center, Shanghai 200052, China

  • Received Date: 2018-12-11
  • Rev Recd Date: 2019-04-03
  • Objective To study the regulation and mechanism of hsa-miRNA-30a-5p on the proliferation of human lung cancer A549 cells. Methods Five pairs of lung cancer and para-carcinoma tissues were harvested in clinical and measured for hsa-miRNA-30a-5p and SCARA5 levels by real-time PCR and western blotting,respectively.The theoretical binding site of hsa-miRNA-30a-5p in SCARA5's 3'UTR was predicted by bioinformatics,and validated by luciferase report assay.A549 cells were transfected with pshRNA-SCARA5 and miRNA-30a-5p inhibitor and 48 h later,the proliferation of A549 cells after genetic intervention was assayed by the MTT method,and the cell growth curve was plotted to observe the effect of hsa-miRNA-30a-5p knockdown on cell proliferation. Results For all five pairs of samples tested,hsa-miRNA-30a-5p was higher in the cancer tissues than in the adjacent tissue(P<0.05),and SCARA5 protein was lower in the cancer tissues than in the adjacent tissue (P<0.05).The bioinformatics analysis showed that there was a theoretical binding site of hsa-miRNA-30a-5p in SCARA5's 3'UTR.The luciferase assay showed that miRNA-30a mimics inhibited the luciferase expressed by the luciferase reporter vector carrying a wild-type SCARA5's 3'UTR(pGL3-WT- SCARA5)(P<0.05,vs pGL3-WT-SCARA5 alone),and miRNA-30a-5p inhibitor enhanced the luciferase activity(P<0.05,vs pGL3-WT- SCARA5 alone).No change was observed when miRNA-30a-5p mimics or miRNA-30a-5p inhibitor was co-transfected with the luciferase reporter vector carrying a mutant SCARA5's 3'UTR(pGL3-MT- SCARA5)(P>0.05,vs pGL3-WT- SCARA5 alone).Hsa-miRNA-30a-5p level in A549 cells transfected with miRNA-30a inhibitor for 48 h was significantly decreased(P<0.05,vs cell control or NC),and there was no difference in hsa-miRNA-30a-5p between the negative control and the transfection control(P>0.05).The proliferation of A549 cells was suppressed by transfection with miRNA-30a-5p inhibitor 24-72 h after transfection(P<0.05,vs control or NC). Conclusion Hsa-miRNA-30a-5p could increase the proliferation in A549 cells via suppressing the expression of SCARA5,and transfection of miRNA-30a-5p inhibitor could inhibit the proliferation of A549 cells via up-regulating SCARA5 expression.
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Study on the mechanism of proliferation of human lung cancer A549 cells regulated by hsa-miRNA-30a-5p

doi: 10.3969/j.issn.1006-0111.2019.05.009
  • 1. Department of Pharmacy, Ningbo Women's and Children's Hospital, Ningbo, 315012, China
  • 2. Hongkou Branch, Changhai Hospital Affiliated to Naval Medical University, Shanghai, 200081, China
  • 3. Department of Pharmacy, Naval Special Medical Center, Shanghai 200052, China
  • Received Date: 2018-12-11
  • Rev Recd Date: 2019-04-03

Abstract: Objective To study the regulation and mechanism of hsa-miRNA-30a-5p on the proliferation of human lung cancer A549 cells. Methods Five pairs of lung cancer and para-carcinoma tissues were harvested in clinical and measured for hsa-miRNA-30a-5p and SCARA5 levels by real-time PCR and western blotting,respectively.The theoretical binding site of hsa-miRNA-30a-5p in SCARA5's 3'UTR was predicted by bioinformatics,and validated by luciferase report assay.A549 cells were transfected with pshRNA-SCARA5 and miRNA-30a-5p inhibitor and 48 h later,the proliferation of A549 cells after genetic intervention was assayed by the MTT method,and the cell growth curve was plotted to observe the effect of hsa-miRNA-30a-5p knockdown on cell proliferation. Results For all five pairs of samples tested,hsa-miRNA-30a-5p was higher in the cancer tissues than in the adjacent tissue(P<0.05),and SCARA5 protein was lower in the cancer tissues than in the adjacent tissue (P<0.05).The bioinformatics analysis showed that there was a theoretical binding site of hsa-miRNA-30a-5p in SCARA5's 3'UTR.The luciferase assay showed that miRNA-30a mimics inhibited the luciferase expressed by the luciferase reporter vector carrying a wild-type SCARA5's 3'UTR(pGL3-WT- SCARA5)(P<0.05,vs pGL3-WT-SCARA5 alone),and miRNA-30a-5p inhibitor enhanced the luciferase activity(P<0.05,vs pGL3-WT- SCARA5 alone).No change was observed when miRNA-30a-5p mimics or miRNA-30a-5p inhibitor was co-transfected with the luciferase reporter vector carrying a mutant SCARA5's 3'UTR(pGL3-MT- SCARA5)(P>0.05,vs pGL3-WT- SCARA5 alone).Hsa-miRNA-30a-5p level in A549 cells transfected with miRNA-30a inhibitor for 48 h was significantly decreased(P<0.05,vs cell control or NC),and there was no difference in hsa-miRNA-30a-5p between the negative control and the transfection control(P>0.05).The proliferation of A549 cells was suppressed by transfection with miRNA-30a-5p inhibitor 24-72 h after transfection(P<0.05,vs control or NC). Conclusion Hsa-miRNA-30a-5p could increase the proliferation in A549 cells via suppressing the expression of SCARA5,and transfection of miRNA-30a-5p inhibitor could inhibit the proliferation of A549 cells via up-regulating SCARA5 expression.

ZHOU Jing, WU Gao, MA Fujia. Study on the mechanism of proliferation of human lung cancer A549 cells regulated by hsa-miRNA-30a-5p[J]. Journal of Pharmaceutical Practice and Service, 2019, 37(5): 433-439. doi: 10.3969/j.issn.1006-0111.2019.05.009
Citation: ZHOU Jing, WU Gao, MA Fujia. Study on the mechanism of proliferation of human lung cancer A549 cells regulated by hsa-miRNA-30a-5p[J]. Journal of Pharmaceutical Practice and Service, 2019, 37(5): 433-439. doi: 10.3969/j.issn.1006-0111.2019.05.009
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