Cargando…
The influence of lentivirus-mediated CXCR4 RNA interference on hepatic metastasis of colorectal cancer
The aim of this study was to construct a lentiviral vector of CXCR4-siRNA (Lenti-CXCR4-siRNA) and investigate whether the vector can inhibit the growth, migration, invasion and hepatic metastasis of colorectal cancer (CRC). RT-PCR and western blotting were employed to identify the ideal RNA interfer...
Autores principales: | , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
D.A. Spandidos
2014
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4063541/ https://www.ncbi.nlm.nih.gov/pubmed/24647809 http://dx.doi.org/10.3892/ijo.2014.2348 |
_version_ | 1782321807464857600 |
---|---|
author | WANG, TIAN-BAO HU, BAO-GUANG LIU, DA-WEI SHI, HAN-PING DONG, WEN-GUANG |
author_facet | WANG, TIAN-BAO HU, BAO-GUANG LIU, DA-WEI SHI, HAN-PING DONG, WEN-GUANG |
author_sort | WANG, TIAN-BAO |
collection | PubMed |
description | The aim of this study was to construct a lentiviral vector of CXCR4-siRNA (Lenti-CXCR4-siRNA) and investigate whether the vector can inhibit the growth, migration, invasion and hepatic metastasis of colorectal cancer (CRC). RT-PCR and western blotting were employed to identify the ideal RNA interference sequence. Lenti-CXCR4-siRNA was constructed and transfected into the SW480 cell line. We used RT-PCR and western blotting to measure the expression of CXCR4 RNA and protein, respectively; the MTS assay to assess the proliferation of SW480 cells; transwell chambers to estimate the inhibitory effect on migration and invasion; and the Balb/c nude mouse model of CRC to examine the inhibition of hepatic metastasis. The relative expression of the CXCR4 gene and protein was 5.4 and 18.95%, respectively, in the siCXCR4 group. The genes in the expression plasmid pLenti-CXCR4-siRNA were in the correct order. In the SW480, nonsense control (NC) and the Lenti-CXCR4-siRNA groups CXCR4 RNA levels were, respectively, 0.54±0.06, 1.00±0.03 and 0.11±0.04 (P=0.0001); CXCR4 protein levels were 0.60±0.03, 0.72±0.03 and 0.18±0.02 (P=0.0001); the OD value was 1.38±0.04 (P=0.0050), 1.28±0.05 (P=0.0256) and 0.92±0.06; SW480 cell number in migration test was 32±6.85, 32.63±1.69 and 0.75±0.71 (P=0.0000); SW480 cell number in the invasion test was 29.13±10.3, 30.38±6.09 and 0.63±0.74 (P=0.0000); hepatic metastasis number was 7.10±3.98 (P=0.034), 7.50±4.09 (P=0.019) and (3.50±2.51); hepatic metastasis mean weight (in g) was 2.25±2.51 (P=0.000), 2.11±2.38 (P=0.000) and 1.45±2.07. Lenti-CXCR4-siRNA constructs were correctly constructed and effectively inhibit the expression of CXCR4 RNA and protein, reducing the proliferation, migration, invasion capacity of SW480 cells and hepatic metastasis of CRC. |
format | Online Article Text |
id | pubmed-4063541 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | D.A. Spandidos |
record_format | MEDLINE/PubMed |
spelling | pubmed-40635412014-06-23 The influence of lentivirus-mediated CXCR4 RNA interference on hepatic metastasis of colorectal cancer WANG, TIAN-BAO HU, BAO-GUANG LIU, DA-WEI SHI, HAN-PING DONG, WEN-GUANG Int J Oncol Articles The aim of this study was to construct a lentiviral vector of CXCR4-siRNA (Lenti-CXCR4-siRNA) and investigate whether the vector can inhibit the growth, migration, invasion and hepatic metastasis of colorectal cancer (CRC). RT-PCR and western blotting were employed to identify the ideal RNA interference sequence. Lenti-CXCR4-siRNA was constructed and transfected into the SW480 cell line. We used RT-PCR and western blotting to measure the expression of CXCR4 RNA and protein, respectively; the MTS assay to assess the proliferation of SW480 cells; transwell chambers to estimate the inhibitory effect on migration and invasion; and the Balb/c nude mouse model of CRC to examine the inhibition of hepatic metastasis. The relative expression of the CXCR4 gene and protein was 5.4 and 18.95%, respectively, in the siCXCR4 group. The genes in the expression plasmid pLenti-CXCR4-siRNA were in the correct order. In the SW480, nonsense control (NC) and the Lenti-CXCR4-siRNA groups CXCR4 RNA levels were, respectively, 0.54±0.06, 1.00±0.03 and 0.11±0.04 (P=0.0001); CXCR4 protein levels were 0.60±0.03, 0.72±0.03 and 0.18±0.02 (P=0.0001); the OD value was 1.38±0.04 (P=0.0050), 1.28±0.05 (P=0.0256) and 0.92±0.06; SW480 cell number in migration test was 32±6.85, 32.63±1.69 and 0.75±0.71 (P=0.0000); SW480 cell number in the invasion test was 29.13±10.3, 30.38±6.09 and 0.63±0.74 (P=0.0000); hepatic metastasis number was 7.10±3.98 (P=0.034), 7.50±4.09 (P=0.019) and (3.50±2.51); hepatic metastasis mean weight (in g) was 2.25±2.51 (P=0.000), 2.11±2.38 (P=0.000) and 1.45±2.07. Lenti-CXCR4-siRNA constructs were correctly constructed and effectively inhibit the expression of CXCR4 RNA and protein, reducing the proliferation, migration, invasion capacity of SW480 cells and hepatic metastasis of CRC. D.A. Spandidos 2014-03-19 /pmc/articles/PMC4063541/ /pubmed/24647809 http://dx.doi.org/10.3892/ijo.2014.2348 Text en Copyright © 2014, Spandidos Publications http://creativecommons.org/licenses/by/3.0 This is an open-access article licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported License. The article may be redistributed, reproduced, and reused for non-commercial purposes, provided the original source is properly cited. |
spellingShingle | Articles WANG, TIAN-BAO HU, BAO-GUANG LIU, DA-WEI SHI, HAN-PING DONG, WEN-GUANG The influence of lentivirus-mediated CXCR4 RNA interference on hepatic metastasis of colorectal cancer |
title | The influence of lentivirus-mediated CXCR4 RNA interference on hepatic metastasis of colorectal cancer |
title_full | The influence of lentivirus-mediated CXCR4 RNA interference on hepatic metastasis of colorectal cancer |
title_fullStr | The influence of lentivirus-mediated CXCR4 RNA interference on hepatic metastasis of colorectal cancer |
title_full_unstemmed | The influence of lentivirus-mediated CXCR4 RNA interference on hepatic metastasis of colorectal cancer |
title_short | The influence of lentivirus-mediated CXCR4 RNA interference on hepatic metastasis of colorectal cancer |
title_sort | influence of lentivirus-mediated cxcr4 rna interference on hepatic metastasis of colorectal cancer |
topic | Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4063541/ https://www.ncbi.nlm.nih.gov/pubmed/24647809 http://dx.doi.org/10.3892/ijo.2014.2348 |
work_keys_str_mv | AT wangtianbao theinfluenceoflentivirusmediatedcxcr4rnainterferenceonhepaticmetastasisofcolorectalcancer AT hubaoguang theinfluenceoflentivirusmediatedcxcr4rnainterferenceonhepaticmetastasisofcolorectalcancer AT liudawei theinfluenceoflentivirusmediatedcxcr4rnainterferenceonhepaticmetastasisofcolorectalcancer AT shihanping theinfluenceoflentivirusmediatedcxcr4rnainterferenceonhepaticmetastasisofcolorectalcancer AT dongwenguang theinfluenceoflentivirusmediatedcxcr4rnainterferenceonhepaticmetastasisofcolorectalcancer AT wangtianbao influenceoflentivirusmediatedcxcr4rnainterferenceonhepaticmetastasisofcolorectalcancer AT hubaoguang influenceoflentivirusmediatedcxcr4rnainterferenceonhepaticmetastasisofcolorectalcancer AT liudawei influenceoflentivirusmediatedcxcr4rnainterferenceonhepaticmetastasisofcolorectalcancer AT shihanping influenceoflentivirusmediatedcxcr4rnainterferenceonhepaticmetastasisofcolorectalcancer AT dongwenguang influenceoflentivirusmediatedcxcr4rnainterferenceonhepaticmetastasisofcolorectalcancer |