[The morphological changes of SW620 cells induced by over-expression of human sorting nexin 3].
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.


CONCLUSIONS
Over-expression of hSNX3 alters the morphology of SW620 cells, but hardly affects cell migration.
Department of Laboratory Medicine, First Affiliated Hospital, Chongqing Medical University, Chongqing 400016, China.
Wei
W
Yang
Medical Research Center, Second Affiliated Hospital, Chengdu Second Clinical College, Chongqing Medical University, Chengdu 610031, China.
Biran
B
Pan
Medical Research Center, Second Affiliated Hospital, Chengdu Second Clinical College, Chongqing Medical University, Chengdu 610031, China.
Tongtong
T
Zhang
Department of Laboratory Medicine, First Affiliated Hospital, Chongqing Medical University, Chongqing 400016, China.
Zhanhao
Z
Wang
Department of Laboratory Medicine, First Affiliated Hospital, Chongqing Medical University, Chongqing 400016, China.
Liping
L
Zhang
Medical Research Center, Second Affiliated Hospital, Chengdu Second Clinical College, Chongqing Medical University, Chengdu 610031, China.
Yuanbiao
Y
Guo
D015153:Blotting, Western; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000235:genetics; D048430:Cell Shape / Q000235:genetics; D003110:Colonic Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D015972:Gene Expression Regulation, Neoplastic; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D008856:Microscopy, Fluorescence; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D058305:Sorting Nexins / Q000235:genetics / Q000378:metabolism*
D011993:Recombinant Fusion Proteins; C513980:SNX3 protein, human; D058305:Sorting Nexins; D049452:Green Fluorescent Proteins