Objective：Early metastasis is a major biological feature of pancreatic cancer.The current study examined whether silencing Slc38a1,a gene involved in energy metabolism,using short hairpin RNA （shRNA） could inhibit ...Objective：Early metastasis is a major biological feature of pancreatic cancer.The current study examined whether silencing Slc38a1,a gene involved in energy metabolism,using short hairpin RNA （shRNA） could inhibit the growth,migration,and invasiveness of pancreatic cancer cells.Methods：A series of Slc38a1 shRNAs were designed and cloned into the pGPU6/GFP/Neo vectors.An shRNA with the most efficacious inhibitory action on SCL38A1 expression （65％ inhibition） upon screening in DH5α bacteria was used to transfect SW1990 human pancreatic cancer cells.Cell growth,migration,and invasiveness were examined using cell counting kit-8,Boyden chamber without and with Matrigel,respectively.Results：Transfection of SW1990 cells with the SLCs38A1 shRNA significantly decreased the proliferation （P＜0.0001） and migratory potential （by 46.7％,P=0.0399） of the cancer cells.Invasiveness,however,was not affected.Conclusions：Inhibiting Slc38a1 using shRNA technology could decrease the growth and migration of representative pancreatic cancer cells.However,the fact that invasiveness was not affected suggested that SLC38A1 is unlikely to be responsible for early metastasis.展开更多
Current studies have demonstrated that SLC38A1 proteins play a causal role in neoplastic cell transformation. The twofold aim of this study was to provide insight into whether a variance in the expression of SLC38A1 e...Current studies have demonstrated that SLC38A1 proteins play a causal role in neoplastic cell transformation. The twofold aim of this study was to provide insight into whether a variance in the expression of SLC38A1 exists between human colorectal cancer and healthy human tissues and to determine how silencing or overexpressing the SLC38A1 gene could affect the proliferation, viability and migration of colorectal cancer cells. Immunohistochemical staining was used to analyze the expression of SLC38A1 in colorectal cancer tissues and adjacent normal mucosa in 77 patients who underwent surgical resection. The expression of SLC38A1 in colorectal cancer tissues and cell lines was detected using RT-PCR and Western blotting. Two colorectal cancer cell lines SW480 and HCT116 were used to examine whether silencing SLC38A1 with si RNA and overexpressing SLC38A1 with sh RNA could affect cell viability and migration. As a result, the SLC38A1 protein was very low or undetectable in the normal colon mucosa. In contrast, strong staining of SLC38A1 protein was found in the cytoplasm in 79.2% colorectal cancer samples. More pronounced SLC38A1 expression in colorectal cancer tissues was significantly associated with tumor node metastasis（TNM） stage. Inhibition of SLC38A1 reduced tumour growth and suppressed proliferation and migration of SW480 cells. In contrast, overexpression of SLC38A1 had the opposite effects on HCT116 cells. SLC38A1 is overexpressed in colorectal cancer, which suggests that it is associated with tumour progression. These results encourage the exploration of SLC38A1 as a target for intervention in colorectal cancer.展开更多
文摘Objective：Early metastasis is a major biological feature of pancreatic cancer.The current study examined whether silencing Slc38a1,a gene involved in energy metabolism,using short hairpin RNA （shRNA） could inhibit the growth,migration,and invasiveness of pancreatic cancer cells.Methods：A series of Slc38a1 shRNAs were designed and cloned into the pGPU6/GFP/Neo vectors.An shRNA with the most efficacious inhibitory action on SCL38A1 expression （65％ inhibition） upon screening in DH5α bacteria was used to transfect SW1990 human pancreatic cancer cells.Cell growth,migration,and invasiveness were examined using cell counting kit-8,Boyden chamber without and with Matrigel,respectively.Results：Transfection of SW1990 cells with the SLCs38A1 shRNA significantly decreased the proliferation （P＜0.0001） and migratory potential （by 46.7％,P=0.0399） of the cancer cells.Invasiveness,however,was not affected.Conclusions：Inhibiting Slc38a1 using shRNA technology could decrease the growth and migration of representative pancreatic cancer cells.However,the fact that invasiveness was not affected suggested that SLC38A1 is unlikely to be responsible for early metastasis.
基金supported in part by grants from National Natural Science Foundation of China(No.81072152)Research Foundation of Health and Family Planning Commission of Hubei Province(No.WJ2015MA010)+1 种基金Natural Science Foundation of Hubei Province(No.2015CFA027)Clinical Medical Research Center of Peritoneal Cancer of Wuhan(No.2015060911020462)
文摘Current studies have demonstrated that SLC38A1 proteins play a causal role in neoplastic cell transformation. The twofold aim of this study was to provide insight into whether a variance in the expression of SLC38A1 exists between human colorectal cancer and healthy human tissues and to determine how silencing or overexpressing the SLC38A1 gene could affect the proliferation, viability and migration of colorectal cancer cells. Immunohistochemical staining was used to analyze the expression of SLC38A1 in colorectal cancer tissues and adjacent normal mucosa in 77 patients who underwent surgical resection. The expression of SLC38A1 in colorectal cancer tissues and cell lines was detected using RT-PCR and Western blotting. Two colorectal cancer cell lines SW480 and HCT116 were used to examine whether silencing SLC38A1 with si RNA and overexpressing SLC38A1 with sh RNA could affect cell viability and migration. As a result, the SLC38A1 protein was very low or undetectable in the normal colon mucosa. In contrast, strong staining of SLC38A1 protein was found in the cytoplasm in 79.2% colorectal cancer samples. More pronounced SLC38A1 expression in colorectal cancer tissues was significantly associated with tumor node metastasis（TNM） stage. Inhibition of SLC38A1 reduced tumour growth and suppressed proliferation and migration of SW480 cells. In contrast, overexpression of SLC38A1 had the opposite effects on HCT116 cells. SLC38A1 is overexpressed in colorectal cancer, which suggests that it is associated with tumour progression. These results encourage the exploration of SLC38A1 as a target for intervention in colorectal cancer.
