AIM： TO identify differentially expressed microRNAs （miRNAs） in human colon cancer stem cells （SW1116csc） and study their function in SW1116csc proliferation. METHODS： SW1116csc were isolated from the human colo...AIM： TO identify differentially expressed microRNAs （miRNAs） in human colon cancer stem cells （SW1116csc） and study their function in SW1116csc proliferation. METHODS： SW1116csc were isolated from the human colon cancer cell line, SW1116 and cultured in serum- free medium. A miRNA microarray was used to detect differential expression profiles of rniRNAs in SW1116csc and SW1116 cells. Real-time quantitative polymerase chain reaction （PCR） was performed to verify the dif- ferential expression of candidate miRNAs obtained from the microarray. Target mRNAs of differentially expressed miRNAs were predicted with target predic- tion tools, miRNA expression plasmids were transfected into SW1116csc using Lipofectamine 2000 reagent. Cell proliferation curves were generated with trypan blue staining, and the colony formation rate of transfected cells was measured with the soft agar colony formation assay. Expression of target mRNAs and proteins from differentially expressed miRNAs were detected using reverse transcription （RT）-PCR and western blotting.RESULTS： Compared with expression in SW1116 cells, 35 miRNAs （including hsa-miR-192, hsa-miR-29b, hsa-miR-215, hsa-miR-194, hsa-miR-33a and hsa- miR-32） were upregulated more than 1.5-fold, and 11 miRNAs （including hsa-miR-93, hsa-miR-1231, hsa- miRPlus-F1080, hsa-miR-524-3p, hsa-miR-886-3p and hsa-miR-561） were downregulated in SW1116csc. The miRNA microarray results were further validated with quantitative RT-PCR. miR-93 was downregulated, and its predicted mRNA targets included BAMBI, CCND2, CDKNIA, HDACS, KIF23, MAP3K9, MAP3K11, MYCN, PPARD, TLE4 and ZDHHCl. Overexpressed miR-93 sig- nificantly inhibited cell proliferation and colony forma- tion by SW1116csc. Furthermore, miR-93 negatively regulated the mRNA and protein levels of HDAC8 and TLE4. CONCLUSION： Some miRNAs were differentially ex- pressed during differentiation of SW1116csc into SW1116 cells, miR-93 may inhibit SW1116csc proliferation and colony formation.展开更多
Mesenchymal stem cells(MSCs)capable of tumour topotaxis have been served as cellular vehicles to deliver anti-tumour agents.As cellular components of the tumour microenvironment,MSCs also affect tumour progression.How...Mesenchymal stem cells(MSCs)capable of tumour topotaxis have been served as cellular vehicles to deliver anti-tumour agents.As cellular components of the tumour microenvironment,MSCs also affect tumour progression.However,the tumour transformation-related genes of MSCs remain unclear since either tumorigenic or tumour suppressor effects within these cells have been researched.Hence,we aimed to identify potential biomarkers indicative of tumorigenic risk by RNA-seq analysis of human placenta tissue-derived MSCs(hPTMSCs)exposed to the carcinogenic agent,3-methylcholanthrene(3-MC).Twenty-nine tumour transformation-related genes and three pluripotency-related genes were appraised as differentially expressed genes(DEGs)in hPTMSCs.Overexpression of sfrp1 led to reduced cell viability,migration,and colony formation in A549.In contrast,the overexpression of ptgs2 exerted the opposite effect.These results indicate that A549 cells with high ptgs2 expression but low sfrp1 expression may have a more potential tumorigenic capacity.Taken together,this study suggests that ptgs2 and sfrp1 may be tumorigenic risk genes.展开更多
In our previous study, five homologous feeder cell lines, Monkey ear skin fibroblasts （MESFs）, clonally derived fibroblasts from the MESFs （CMESFs）, monkey oviductal fibroblasts （MOFs）, monkey follicular granulo...In our previous study, five homologous feeder cell lines, Monkey ear skin fibroblasts （MESFs）, clonally derived fibroblasts from the MESFs （CMESFs）, monkey oviductal fibroblasts （MOFs）, monkey follicular granulosa fibroblast-like （MFGs） cells, monkey follicular granulosa epithelium-like （MFGEs） cells, were developed for the maintenance of rhesus embryonic stem cells （rESCs）. We found that MESFs, CMESFs, MOFs and MFGs, but not MFGEs, support the growth of rhesus embryonic stem cells. Moreover, we detected some genes that are upregulated in supportive feeder cell lines by semi-quantitative PCR. In the present study, we applied the GeneChip Rhesus Macaque Genome Array of Affymetrix Corporation to study the expression profiles of these five feeder cell lines, in purpose to find out which cytokines and signaling pathways were important in maintaining the rESCs, mRNAs of eight genes, including GREM2, bFGF, KITLG, DKK3, GREM1, AREG, SERPINF1 and LTBP1, were found to be upregulated in supportive feeder cell lines, but not in MFGE. The results indicate that many signaling pathways may play redundant roles in supporting the undifferentiated growth and maintenance of pluripotency in rESCs.展开更多
基金Supported by Medical guidance projects of Shanghai Science Committee,No.10411961800Youth Science Foundation of Fudan University,No.08FQ49
文摘AIM： TO identify differentially expressed microRNAs （miRNAs） in human colon cancer stem cells （SW1116csc） and study their function in SW1116csc proliferation. METHODS： SW1116csc were isolated from the human colon cancer cell line, SW1116 and cultured in serum- free medium. A miRNA microarray was used to detect differential expression profiles of rniRNAs in SW1116csc and SW1116 cells. Real-time quantitative polymerase chain reaction （PCR） was performed to verify the dif- ferential expression of candidate miRNAs obtained from the microarray. Target mRNAs of differentially expressed miRNAs were predicted with target predic- tion tools, miRNA expression plasmids were transfected into SW1116csc using Lipofectamine 2000 reagent. Cell proliferation curves were generated with trypan blue staining, and the colony formation rate of transfected cells was measured with the soft agar colony formation assay. Expression of target mRNAs and proteins from differentially expressed miRNAs were detected using reverse transcription （RT）-PCR and western blotting.RESULTS： Compared with expression in SW1116 cells, 35 miRNAs （including hsa-miR-192, hsa-miR-29b, hsa-miR-215, hsa-miR-194, hsa-miR-33a and hsa- miR-32） were upregulated more than 1.5-fold, and 11 miRNAs （including hsa-miR-93, hsa-miR-1231, hsa- miRPlus-F1080, hsa-miR-524-3p, hsa-miR-886-3p and hsa-miR-561） were downregulated in SW1116csc. The miRNA microarray results were further validated with quantitative RT-PCR. miR-93 was downregulated, and its predicted mRNA targets included BAMBI, CCND2, CDKNIA, HDACS, KIF23, MAP3K9, MAP3K11, MYCN, PPARD, TLE4 and ZDHHCl. Overexpressed miR-93 sig- nificantly inhibited cell proliferation and colony forma- tion by SW1116csc. Furthermore, miR-93 negatively regulated the mRNA and protein levels of HDAC8 and TLE4. CONCLUSION： Some miRNAs were differentially ex- pressed during differentiation of SW1116csc into SW1116 cells, miR-93 may inhibit SW1116csc proliferation and colony formation.
基金supported by grants from the Ningxia Science and Technological Supporting Project(2015KJHM38)the Ningxia Natural Science Foundation(2019AAC03231).
文摘Mesenchymal stem cells(MSCs)capable of tumour topotaxis have been served as cellular vehicles to deliver anti-tumour agents.As cellular components of the tumour microenvironment,MSCs also affect tumour progression.However,the tumour transformation-related genes of MSCs remain unclear since either tumorigenic or tumour suppressor effects within these cells have been researched.Hence,we aimed to identify potential biomarkers indicative of tumorigenic risk by RNA-seq analysis of human placenta tissue-derived MSCs(hPTMSCs)exposed to the carcinogenic agent,3-methylcholanthrene(3-MC).Twenty-nine tumour transformation-related genes and three pluripotency-related genes were appraised as differentially expressed genes(DEGs)in hPTMSCs.Overexpression of sfrp1 led to reduced cell viability,migration,and colony formation in A549.In contrast,the overexpression of ptgs2 exerted the opposite effect.These results indicate that A549 cells with high ptgs2 expression but low sfrp1 expression may have a more potential tumorigenic capacity.Taken together,this study suggests that ptgs2 and sfrp1 may be tumorigenic risk genes.
文摘In our previous study, five homologous feeder cell lines, Monkey ear skin fibroblasts （MESFs）, clonally derived fibroblasts from the MESFs （CMESFs）, monkey oviductal fibroblasts （MOFs）, monkey follicular granulosa fibroblast-like （MFGs） cells, monkey follicular granulosa epithelium-like （MFGEs） cells, were developed for the maintenance of rhesus embryonic stem cells （rESCs）. We found that MESFs, CMESFs, MOFs and MFGs, but not MFGEs, support the growth of rhesus embryonic stem cells. Moreover, we detected some genes that are upregulated in supportive feeder cell lines by semi-quantitative PCR. In the present study, we applied the GeneChip Rhesus Macaque Genome Array of Affymetrix Corporation to study the expression profiles of these five feeder cell lines, in purpose to find out which cytokines and signaling pathways were important in maintaining the rESCs, mRNAs of eight genes, including GREM2, bFGF, KITLG, DKK3, GREM1, AREG, SERPINF1 and LTBP1, were found to be upregulated in supportive feeder cell lines, but not in MFGE. The results indicate that many signaling pathways may play redundant roles in supporting the undifferentiated growth and maintenance of pluripotency in rESCs.
