乳腺癌;α2,8-唾液酸转移酶4;侵袭;迁移," /> 乳腺癌;α2,8-唾液酸转移酶4;侵袭;迁移,"/> ;α2, 8-sialyltransferase 4InvasionMigration,"/> <span>Effects of shRNA silencing ST8SIA4 gene expression on invasion and migration of breast cancer cells</span>

Chinese Clinical Oncology ›› 2018, Vol. 23 ›› Issue (10): 870-874.

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Effects of shRNA silencing ST8SIA4 gene expression on invasion and migration of breast cancer cells

  

  1. the Fourth Department of Breast Cancerthe Affiliated Zhongshan Hospital of Dalian UniversityDalian 116001
  • Received:2018-03-22 Revised:2018-08-19 Online:2018-10-31 Published:2019-03-20

Abstract: Objective  To investigate the effect of short hairpin RNA shRNA targeted silencing of α2 8-sialyltransferase 4 ST8SIA4 gene on invasion and migration of breast cancer BT549 cells. Methods  Real-time quantitative PCR QPCR and Western blotting were used to detect the mRNA and protein levels of ST8SIA4 in BT549 cells and MCF-10A cells. BT549 cells were randomly transfected with the successfully constructed shRNA vector fragments targeting ST8SIA4 gene silencing group or nonsense shRNA fragments control group via LipofectamineTM 2000. ST8SIA4 mRNA expression was detected after 48 h transfection by QPCR. The expressions of ST8SIA4 phosphorylated Akt p-Akt), neuropilin2 NRP2 and tumor necrosis factor-α TNF-α) were detected by Western blotting. The invasion and migration abilities of cells were detected by Transwell assay and scratch assay respectively. Results Compared with MCF-10A cells the relative expression level of ST8SIA4 mRNA and protein in BT549 cells increased significantly P0.05. After 48 h transfection the mRNA and protein levels of ST8SIA4 in the silencing group were 0.19±0.02 and 0.13±0.02 lower than 0.98±0.11 and 0.52±0.05 in the control group P0.05. The number of invasive cells migration rate and relative expressions of p-Akt NRP2 and TNF-α were 39.5±4.2 16.9±1.3)%, 0.22±0.03 0.31±0.04 and 0.27±0.06 in the silencing group all lower than 91.3±8.5 38.4±3.9)%, 0.58±0.07 0.55±0.05 and 0.46±0.05 in the control groupP0.05. Conclusion  ST8SIA4 gene is highly expressed in breast cancer BT549 cells. Silencing its expression may inhibit invasion and migration which may be related to up-regulation of p-Akt NRP2 and TNF-a protein expression.

Key words: ;α2')"> Breast cancer;α2, -sialyltransferase 4InvasionMigration')"> 8-sialyltransferase 4InvasionMigration

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