Wei Yang1, Biran Pan2, Tongtong Zhang2, Zhanhao Wang1, Liping Zhang1, Yuanbiao Guo2. 1. Department of Laboratory Medicine, First Affiliated Hospital, Chongqing Medical University, Chongqing 400016, China. 2. Medical Research Center, Second Affiliated Hospital, Chengdu Second Clinical College, Chongqing Medical University, Chengdu 610031, China.
Abstract
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.
OBJECTIVE: To explore the effect of humansorting 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.