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[The morphological changes of SW620 cells induced by over-expression of human sorting nexin 3]
Wei Yang1, Biran Pan2, Tongtong Zhang2
1Department of Laboratory Medicine, First Affiliated Hospital, Chongqing Medical University, Chongqing 400016, China.
Objective:
To explore the effect of human sorting nexin 3 (hSNX3) on the morphology and migration of colorectal cancer cells through over-expression of hSNX3 in SW620 cells.
Methods:
A lentiviral vector pEZ-Lv201 (pEZ-SV40-eGFP-IRES-Puro) inserted with hSNX3 gene was transferred into SW620 cells to construct and package recombinant lentiviral expression vector pEZ-hSNX3-Lv201. The cell clones stably over-expressing hSNX3 (SW620hSNX3) were screened to observe cell morphology and migration ability using Transwell(TM) assay and wound scratch assay. E-cadherin was detected by Western blotting.
Results:
Sequence analysis demonstrated that the lentiviral expression vector pEZ-hSNX3-Lv201 was successfully constructed. The packaging titers of the recombinant lentivirus and control virus were 2.12×10(9) copies/mL and 7.9×10(10) copies/mL, respectively. After SW620 cells were transfected with the lentivirus, we got stable hSNX3-overexpressed SW620hSNX3 cells. Most of SW620hSNX3 cells (88%) became oval instead of the original spindle-like shape. However, cell migration ability and expression of E-cadherin showed no significant differences between SW620hSNX3 cells and SW620 cells.
Conclusion:
Over-expression of hSNX3 alters the morphology of SW620 cells, but hardly affects cell migration.
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