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Objective To understand the current status of infections of intestinal parasites of population in Taixing City,Ji-angsu Province. Methods The infection rates and densities of human intestinal parasites were investigated according to the methods of the National Investigation Scheme on Human Principal Parasites,and the data of society,economy and disease con-trol were collected and analyzed. Results Among 2 556 people investigated in five villages,16 persons were found with intesti-nal parasites, with an infection rate of 0.63%. The infection rate was higher in residents with a low education level than in others and it was higher in the age group over 50 years than in the group under 50 years. The infection density was mild and the most was the single parasite infection. Conclusions The current status of intestinal parasite infections of population in Taixing City has reached the county-level control standard of Jiangsu Province. Therefore,the preventive strategy and measures should be ad-justed and the monitoring work should be strengthened.
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Objective To investigate the effects of a proliferation-inducing ligand(APRIL) on migration and invasion of colorectal cancer (CRC) and matrix metalloproteinases (MMPs) expression in order to observe the role of APRIL in CRC metastasis.Methods The siRNA plasmid vector targeting APRIL gene (siRNA-APRIL) was transfected into SW480 cells and recombinant human APRIL(rhAPRIL) was used to stimulate HCT-116 cells.Tumor cell migration and invasion were measured by Transwell chambers.RT-PCR and ELISA were applied to examine the expression level of MMPs.Results Metastatic and invasive capacities of siRNA-APRIL transfected SW480 were significantly inhibited,and these capacities of APRIL stimulated HCT-116 cells were significantly enhanced compared with their respective controls( all P＜0.05 ),accompanied with the alterations of MMPs mRNA and secreted protein expression( P＜0.05).The number of invading cells of SW480 control and rhAPRIL stimulated HCT-116 was significantly decreased by a MMP inhibitor GM6001 ( P＜0.05 ).Conclusion APRIL facilitates migration and invasion of CRC via regulation of MMPs,which suggests that APRIL might be used as a new target for the intervention and treatment of CRC metastasis.
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Objective To prove the remarkable inhibitive effect of multiple siRNAs targeting a proliferation-inducing ligand (APRIL) on the human colorectal cancer cell.Methods We constructed a multiple short hairpin RNA(shRNA) expression vector containing four shRNAs (pG4) as well as four single one (pGsh644,pGsh1451,pGsh1938,pGsh2231) against APRIL gene in SW480 cell,and then transfected them into the human colorectal cancer cell line by cationic liposome.Ultimately,SW480 were screened by EGFP to obtain expression cell lines.APRIL expression levels including mRNA and APRIL protein were detected after transfected with all different kinds of vectors.Results A multiple shRNA expression vector containing four shRNAs (pG4) and four single ones were successfully constructed.Four single vectors (pGsh644,pGsh1451,pGsh1938,pGsh2231) and the multiple siRNAs expression vector (pG4) all decreased the APRIL mRNA by 56.2％,49.5％ ;50.9％,49.2％ and 79.3％.And APRIL protein expression was also remarkably reduced,especially by multiple siRNAs expression vector(87.5％).Conclusion Multiple siRNAs expression vector produced a more significant knockdown effect of APRIL than the vectors containing only one APRIL shRNA.What we found suggested us using the vector containing multiple shRNA to silence the expression of APRIL might be exploited as a novel therapeutic strategy for tumors.
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Objective To construct and screen siRNA targeting a proliferation-inducing ligand (APRIL) gene in a mouse colorectal cancer celline, CT-26. To investigate the effects to the cell growth and migrant capacity of CT-26 after knockdown APRIL gene, lay the foundation for molecular targeted therapy to colorectal cancer. Methods Four pairs of APRIL siRNA were designed and chemically synthesized. And disorder sequences were synthesized as a negative control. These sequences were transfected with LipofectAMINE 2000 into CT-26 cells, which high-expressed APRIL gene. The transfection efficency rate of 6-FAM labelled control siRNA was detected by fluorescence microscope. The inhibition effectiveness of APRIL mRNA and protein was analyzed by FQ-RT-PCR and Western blot, respectively. Cell proliferation activity was analyzed by cell counting kit-8, cell migration capacity was detected by the repair of cell damage, and MMP-2 together with TIMP-1, two important regulatory genes in cell metastasis, were measured by RT-PCR.Results The different kinds of APRIL siRNA effectively suppressed the level of APRIL mRNA and the protein expression in CT-26 (P ＜ 0.05 ). Cell proliferation and metastasis ability were repressed after APRIL siRNA transfection( P ＜ 0.05 ), compared with random siRNA control and nontransfected control. The mRNA levels of MMP-2 and TIMP-1 genes wre significantly altered among APRIL siRNA groups and two control groups ( P ＜ 0.05). Conclusion We have constructed and screened a kind of siRNA (APsi737) targeting APRIL gene in a mouse colorectal cancer cell line, CT-26. APRIL siRNA can effectively inhibit the cell growth and migration capacity, maybe be regulated by MMP-2 and TIMP-1.
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Objective To investigate the effects of a proliferation-inducing ligand(APRIL)gene silencing by small interfering RNA(siRNA)on cell cycle and proliferation of colon carcinoma SW480 cells.Methods The siRNA plasmid vector targeting APRIL gene,named as siRNA-APRIL,was transfected into SW480 cells,transfected with scrambled vector as a nontargeting control and nontransfected group as another control.APRIL mRNA and protein expression were examined by real-time PCR and Western blot,respectively.Cell proliferation activity was analyzed by cell counting kit-8(CK-8),cell cycle was detected by flow cytometry,and p21 together with p27,two important regulatory genes in cell cycle,were measured by RTPCR.Results Compared with nontargeting control and nontransfected control,APRIL expression was inhibited significantly at both mRNA and protein level by siRNA-APRIL being transfected in SW480 cells(P ＜0.05).Cell proliferation ability was drastically repressed after siRNA-APRIL being transfected at 48 h,72 h and 96 h(P ＜ 0.05).After transfected 48 h,the percent of Go/G1 phase cell was significantly increased,S and G2/M phase cell were significantly decreased,the number of cell in apoptosis was increased and the expression of p21 and p27 mRNA were up-regulated(P ＜ 0.05).There was no significant difference when compared the two control groups each other(P ＞ 0.05).Conclusion siRNA-APRIL can effectively knockdown the expression of APRIL gene in SW480 cells,moreover,it can inhibit the cell proliferation and induce G0/G1 phase cell cycle arrest,which occurrence may involve in upregulation the mRNA expression of p21 and p27.
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Objective To study the influence of short hairpin RNA-mediated inhibition of APRIL gene on proliferation,invasion and metastasis of human colon carcinoma SW480 ceils in vitro. Methods After treatment with shRNA, the expression level of APRIL protein in human colon carcinoma SW480 cells was detected by western blotting. And cell adhesion, cell migration and cell invasion of SW480 cells transfected with sh637 were analyzed respectively as compared with non-transfected SW480 cells and mocktransfectant. Results The expression of APRIL protein in SW480 transfected with sh637 were significantly lower than mock-plasmid groups and untransfected ones (F=42.15, P=0.00). Cell proliferation was markedly inhibited, compared with untransfected groups and negative control ones (F=24.76, P<0.05).And the average absorption of cell adhesion in transfected groups, mock-plasmid and non-transfected ones were 0.343, 0.409, 0.412, respectively. Cell adhesion decreased 39% compared with untransfected groups. Similarly, there was significant difference for cell migration (F=65.53, P<0.01) between transfected groups (The average ceil number in exposed area to migrate was 47.89±13.16) and nontransfected (98.78±23.26) as well as mock-plasmid ones (108.01±39.11). Then, in view of the average absorption of cell invasion between transfected groups(0.58±0.10) and non-transfected (0.94±0.23) as well as mock-plasmid ones(1.11±0.21), a prominently difference for cell invasion was also found between them (F=5.771, P<0.05). Conclusion The results suggest APRIL has a proliferation-inducing effect and probably be involved in cell invasion and cell metastasis of colon carcinoma and it could enhance capacity of invasion and metastasis of SW480 cells.