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Immature hematopoietic progenitors are a constant source for renewal of hemocyte populations and the basic component of the tissue and cell repair apparatus. A unique property of these cells of internalizing extracellular double-stranded DNA has been previously shown. The leukostimulatory effect demonstrated in our pioneering studies was considered to be due to the feature of this cell. In the present research, we have analyzed the effects of DNA genome reconstructor preparation (DNAgr), DNAmix, and human recombinant angiogenin on both hematopoietic stem cells and multipotent progenitors. Treatment with bone marrow cells of experimental mice with these preparations stimulates colony formation by hematopoietic stem cells and proliferation of multipotent descendants. The main lineage responsible for this is the granulocyte-macrophage hematopoietic lineage. Using fluorescent microscopy as well as FACS assay, co-localization of primitive c-Kit- and Sca-1-positive progenitors and the TAMRA-labeled double-stranded DNA has been shown. Human recombinant angiogenin was used as a reference agent. Cells with specific markers were quantified in intact bone marrow and colonies grown in the presence of inducers. Quantitative analysis revealed that a total of 14,000 fragment copies of 500 bp, which is 0.2% of the haploid genome, can be delivered into early progenitors. Extracellular double-stranded DNA fragments stimulated the colony formation in early hematopoietic progenitors from the bone marrow, which assumed their effect on cells in G0. The observed number of Sca1+/c-Kit+ cells in colonies testifies to the possibility of both symmetrical and asymmetrical division of the initial hematopoietic stem cell and its progeny.
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Abstract Lung cancer is a major cause of cancer-related death worldwide. This study investigated the regulatory effects of the microRNA-99a-5p (miR-99a-5)/VLDLR axis on lung cancer cell sensitivity to chemotherapy and its mechanism. miR-99a-5p and VLDLR expression levels were quantified using RT-qPCR and western blotting, respectively. The IC50 value of cisplatin (DDP) was determined using a CCK-8 assay. Lung cancer cell proliferation and apoptosis were measured using the CCK-8 assay and flow cytometry, respectively. The mRNA expression levels of apoptosis-related factors (Bax, Bcl-2, and Caspase-3) were evaluated using RT-qPCR. The direct relationship between miR-99a-5p and VLDLR was validated using dual-luciferase reporter gene and RIP assays. miR-99a-5p was weakly expressed in DDP-resistant lung cancer cells. Overexpression of miR-99a-5p promoted DDP sensitivity, suppressed proliferation and colony formation, and promoted apoptosis of A549/DDP cells in vitro. Mechanistically, miR-99a-5p restrained VLDLR expression by binding to VLDLR 3'UTR, and miR-99a-5p mediated inhibition of VLDLR regulated the DDP sensitivity, proliferation, and apoptosis of A549/ DDP cells. Overexpression of miR-99a-5p inhibited the growth of A549 cells and increased chemosensitivity of A549 cells to DDP in vivo. In conclusion, miR-99a-5p overexpression promotes sensitivity to DDP and cell apoptosis by downregulating VLDLR expression in A549/ DDP cells.
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Objective:To investigate the effects of silencing HOXA13 gene on the malignant phenotypes of hepatocellular carcinoma cell lines HepG2and QGY-7703,and to provide a new molecular target for the diagnosis and treatment of hepatocellular carcinoma.Methods:The interference recombinant plasmid of plent-U6-GFP/si-HOXA13 was constructed,then it was stably transfected into the HepG2and QGY-7703 cells.The interfering effects of HOXA13 gene were determined by RT-PCR and Western blotting methods.The HepG2and QGY-7703 cells transfected with HOXA13 knockdown were used as experimental group,and the HepG2and QGY-7703 cells transfected with empty plasmid were used as control group.Then the growth speed was examined by MTT assay, the cell doubling time was examined by double time assay,the colony formation ability was examined by colony formation assay,and the cell cycle was examined by flow cytometry.Results:The MTT assay results showed that compared with control group,the growth speeds of HepG2and GY-7703 cells in experimental group were significantly decreased and the cell cycles were changed;the number of HepG2and QGY-7703 cells in G1phase was increased and the number of cells in S phase was decreased.The doubling time of HepG2and QGY-7703 cells in control group and experimental group were(4.59±0.27),(4.93±0.17),(6.02±0.86),and(6.43±0.66)h, and the differences between control group and experimental group were significant(P<0.05).The colony number of HepG2and QGY-7703 cells in control group and experimental group were 264.00 ± 12.62,269.00 ± 4.55, 165.00± 10.61,and 215.00 ± 4.43,and the differences between control group and experimental group were significant(P<0.01).Conclusion:HOXA13 can increase the proliferation,enhance the clone formation,decrease the number of cells at G1phase and increase the number of cells at S phase in the hepatocellular carcinoma HepG2 and QGY-7703 cells;and it may used as a new molecular target for diagnosis and treatment of hepatocellular carcinoma.
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Objetive: To investigate the effects of silencing HOXA13 gene on the malignant phenotypes of hepatocellular carcinoma cell lines HepG2 and QGY-7703, and to provide a new molecular target for the diagnosis and treatment of hepatocellular carcinoma. Methods: The interference recombinant plasmid of plent-U6-GFP/si-HOXA13 was constructed, then it was stably transfected into the HepG2 and QGY-7703 cells. The interfering effects of HOXA13 gene were determined by RT-PCR and Western blotting methods. The HepG2 and QGY-7703 cells transfected with HOXA13 knockdown were used as experimental group, and the HepG2 and QGY-7703 cells transfected with empty plasmid were used as control group. Then the growth speed was examined by MTT assay, the cell doubling time was examined by double time assay, the colony formation ability was examined by colony formation assay, and the cell cycle was examined by flow cytometry. Results: The MTT assay results showed that compared with control group, the growth speeds of HepG2 and GY-7703 cells in experimental group were significantly decreased and the cell cycles were changed; the number of HepG2 and QGY-7703 cells in G1 phase was increased and the number of cells in S phase was decreased. The doubling time of HepG2 and QGY-7703 cells in control group and experimental group were (4.59 ± 0.27), (4.93 ± 0.17), (6.02 ± 0.86), and (6.43 ± 0.66) h, and the differences between control group and experimental group were significant (P<0.05). The colony number of HepG2 and QGY-7703 cells in control group and experimental group were 264.00 ± 12.62, 269.00 ± 4.55, 165.00±10.61, and 215.00 ± 4.43, and the differences between control group and experimental group were significant (P<0.01). Conclusion: HOXA13 can increase the proliferation, enhance the clone formation, decrease the number of cells at G1 phase and increase the number of cells at S phase in the hepatocellular carcinoma HepG2and QGY-7703 cells; and it may used as a new molecular target for diagnosis and treatment of hepatocellular carcinoma.
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Objective:To investigate the function and mechanism of cyclin-dependent kinase 4/6 (CDK4/6) inhibitor PD0332991 on clo-nogenicity of mantle cell lymphoma (MCL) cells. Methods:The effect of PD0332991 on MCL cell cycle distribution was assessed by flow cytometry;Western blot was used to test expression level of Rb protein and phosphorylated Rb protein in MCL after treatment with PD0332991;colony forming assay was performed to test the role of PD0332991 and mitoxantrone and their combination on colo-ny forming activity in MCL. Results:Flow cytometry revealed that PD0332991 can increase G0/G1 phase MCL cells and significantly de-crease S phase cells, leading to G0/G1 cell arrest. Western blot confirmed that PD0332991 exerted no effect on Rb protein expression but suppressed levels of phosphorylated Rb protein. Colony forming assay showed that PD0332991 significantly suppressed colony for-mation and enhanced the effect of mitoxantrone on colony forming activity in MCL. Conclusion:This study revealed that CDK4/6 inhib-itor PD0332991 induced G0/G1 cell arrest and increased the effect of mitoxantrone on MCL clonogenicity by suppressing levels of phosphorylated Rb protein.
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Objective:To investigate the effect of human growth hormone releasing hormone receptor splice variant type 1 (GHRHR SV1) on the proliferation of human liver cancer HepG2 cells,and to clarify the proliferation effect of GHRHR SV1 on the human cancer cells.Methods:The GHRHR SV1 plasmids were transfected into the human HepG2 cells to construct the HepG2-SV1 cell line.HepG2 group(HepG2 cells),HepG2-empty group(HepG2-pcDNA3.0 cell line) and HepG2-SV1 group(HepG2-SV1 cells) were set up.PCR and Western blotting methods were used to identify the HepG2-SV1 cell line;CCK-8 method was used to detect prolifernation rate of cells;colony formation assay was used to detect the colony formation rate of cells;cell wound healing assay was used to evaluate the migration rate of cells.Results:The PCR and Western blotting results showed the HepG2-SV1 cell line expressed GHRHR SV1 steadily.The CCK-8 results showed that the proliferation rate of the HepG2-SV1 cells in HepG2-SV1 group was higher than that of the HepG2-pcDNA3.0 cells in HepG2-empty group(P<0.05).The colony formation assay results showed that the colony formation rate of HepG2-SV1 cells in HepG2-SV1 group was 3.5 times higher than that of the HepG2-pcDNA3.0 cells in HepG2-empty group(P<0.05).The cell wound scratch assay results showed that the migration rate of the HepG2-SV1 cells in HepG2-SV1 group was higher than that of the HepG2-pcDNA3.0 cells in HepG2-empty group(P<0.05).Conclusion:GHRHR SV1 could increase the proliferation of HepG2 cells.
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Objective: To establish a radioresistant model of non-small-cell cancer A549, and to provide the experimental basis for further researchonradioresistance. Methods: The cell number and the dose of radiation therapy were confirmed. The intermittent radiation were used to induce the radioresistant cell model. The morphology of cells were observed with an inverted phase contrast microscope. Colony formation was used to identify the radioresistance of the RA549 cell. Results: RA549 cells were longer and bigger than A549 cells; surviving fraction of RA549 was significantly increased than A549. Conclusion: Intermittent radiation can successfully induce the radioresistant cell model of lung cancer cells.
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Aim To investigate the effect of curcumin on radiosensitivity of radioresistant nasopharyngeal carcinoma cell line CNE-2R and its mechanism.Methods The concentration of curcumin was screened by MTT assay.Dose-survival curves were obtained according to the colony forming test for L-Q matching and multitarget-single hitting matching,while SF2 and the correlation parameters of radiation biology were calculated.The changes of cell cycle in CNE-2R cells caused by curcumin were also tested by flow cytometry(FCM).The differential expression of genes related to cell cycle and DNA damage repair were detected by RT-qPCR.Results CNE-2R cells could not be inhibited by 10 μmol·L-1 curcumin.Dealt with 10 μmol·L-1 curcumin for 24 h,the value of α/β increased to 1 596 from 6.56;the value of SF2 decreased to 0.361 Gy from 1.93 Gy;the value of N decreased to 1.06 from 1.60;the value of D0 decreased to 2.12 from 3.27;the value of Dq decreased to 0.12 from 1.53.FCM showed that the cells in G2 phase had a significant increase and the cells in S phase had a significant decrease after dealt with 10 μmol·L-1 curcumin for 24 h.The expression of CDK4 was significantly up-regulated and GADD45g,BRCA1 were significantly down-regulated.Conclusion Curcumin radiosensitizes nasopharyngeal carcinoma cell line CNE-2R by changing cell cycle and affecting DNA damage repair through regulating the expression of CDK4,GADD45 g and BRCA1.
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Objective To investigate the influence of recombinant neuregulin-1 beta (rhNRG-1β) on neural stem cell proliferation through extracellular regulated protein kinases (ERK) signaling pathway in oxygen and glucose deprivation (OGD) environment.Methods Neural stem cells were obtained from embryonic brain of mice pregnant for 14-17 d,cultured and identified by immunochemical staining through detection of the indicator nestin using the SABC-FITC (POD)double standard kit.Neural stem cells were divided into three experiment groups (OGD group,control group and OGD + rhNRG-1β group).Control group:identified neural stem cells,2 × 107,were cultured for 3 h in the 24-hole culture plate with DMEM/F12 complete culture medium;OGD group:neural stem cells,2 × 107,were cultured in the 24-hole culture plate deprived glucose DMEM/F12 in a wet airtight container (37 ℃ constant temperature),cells were cultured with mixed gas of nitrogen (950 ml/L) and oxygen (50 ml/L) for 1 h,and then the culture medium was replaced with complete culture medium and cultured for 3 h;OGD + rhNRG-1β group:before OGD intervention,100 μg/L rhNRG-1β was given for 3 h.Neurospheres formation:the three groups of stem cells were dispersed into single cells,1 × 106/ml cells were inoculated to culture plates containing cover slips coated with poly lysine,and cultivated for 7 d,and neurospheres formation of the 3 groups of neural stem cells was observed under microscope,which was aimed to record neural stem cells proliferation changes.Colony formation:the three groups of stem cells,vaccinated in 60 mm in a petri dish,2 × 107 in number,were cultivated in complete culture medium for 24 h.The colony formation of the three groups of cells was observed under microscope,and neural stem cells proliferation changes were observed.Western blotting:the change of phosphorylation ERK (pERK) protein of the three groups of stem cells was determined,and the effect of pERK protein expression regulated by rhNRG-1β in mice neural stem cells proliferation through ERK signaling pathway was observed.Results Microscopically the primary cultured stem cells grew in single or in pairs,in a round shape;neural stem cell proliferated in clumps or colony;neural stem cells expressed the specificity of nestin protein markers with fluorescent yellow-green color.The differences in the aspects of the average diameter of neurospheres and neurospheres quantity in the neural stem cells between groups were statistically significant (F =693.66,1 002.09,all P ＜ 0.01),and among them the neuropheres formation of OGD group was significantly suppressed.Formation quantity and average diameter in OGD group [(88.78 ± 7.14) numbers,(62.12 ± 2.52) μm] were significantly lower than those in the control group [(246.34 ± 8.67) numbers,(128.45 ± 2.33) μm] and those in OGD + rhNRG-1β group [(237.87 ± 6.61) numbers,(118.37 ± 2.71) μm,all P ＜ 0.01].The difference of colony formation rate of neural stem cell was statistically significant (F =132.03,P ＜ 0.01),and among them colony formation of OGD group significantly suppressed.Formation rate in OGD group [(11.65 ± 0.94)％] was significantly lower than that in the control group [(33.23 ± 2.93)％] and that in OGD + rhNRG-1β group [(31.42 ± 2.61)％,all P ＜ 0.01].Western blotting showed that the difference of pERK protein expression of neural stem cells between groups was statistically significant (F =63.76,P ＜ 0.01).Relative expression of the pERK protein in OGD group (0.487 ± 0.072) was significantly lower than that in the control group (1.013 ± 0.112) and that in OGD + rh-NRG-1β group (1.752 ± 0.278,all P ＜ 0.01).Conclusion rhNRG-1β preserves neural stem cell proliferation with phosphorylation ERK protein expression up-regulated in oxygen and glucose deprivation environment.
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AIM: To detect the protein expression of protein gene product (PGP9.5) in cervical carcinoma samples, and to explore its relationship with clinicopathological features and prognosis of cervical carcinoma patients.The potential value of PGP9.5-siRNA in the treatment of cervical cancer was preliminarily investigated in vitro.METHODS:The clinical data of cervical cancer patients ( n=180 ) who received surgical treatment in Department of Gynecology of Tianjin Fifth Central Hospital and Tianjin Central Hospital of Gynecology and Obstetrics from January 2008 to June 2015 were retrospectively analyzed.Immunohistochemical staining for PGP9.5 in all pathological specimens from cervical cancer biopsy was performed.The patients were divided into high expression of PGP9.5 group and low expression of PGP9.5 group.The relationship between PGP9.5 expression and clinicopathological parameters, such as age, HPV infection, path-ological grade, tumor diameter, lymph node metastasis, depth of invasion and clinical stage, were analyzed.The overall survival was analyzed by Kaplan-Meier method and the log-rank test.The PGP9.5-siRNA, NC-siRNA, PGP9.5 eukaryotic expression plasmid and empty vector were transfected into the SiHa cells, and the effects of PGP9.5 expression on the abili- ties of colony formation and cell invasion were determined by Western blot, colony formation assay and Transwell experi-ment.RESULTS:The relationships between the expression level of PGP9.5 and patients’ pathological characteristics in-cluding grade, tumor size, lymph node metastasis, depth of invasion, vascular involvement and clinical stage were statisti-cally significant ( P<0.05) .The overall survival rates of 3 and 5 years in PGP9.5 high expression group were significantly lower than those in PGP9.5 low expression group ( P<0.05 ) .Compared with control group, the protein expression of PGP9.5, the number of colony formation and the number of invasive cells in si-PGP9.5 group were significantly decreased. The protein expression of PGP9.5, the number of colony formation and the number of invasive cells in PGP9.5 group were significantly increased.CONCLUSION:Over-expression of PGP9.5 protein indicates poor prognosis of cervical cancer, which may be a good predictor for the prognosis of cervical cancer patients.Inhibition of PGP9.5 expression may be an ef-fective way of gene therapy for cervical cancer.
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Objective To establish an Apak gene stable and permanent knockout cell line using CRISPR/Cas9 system in human colon cancer cells ( HCT116 cells), and study the effect of Apak knock-out on p53 activity and apoptosis. Methods The lentiCRISPR v2-sgRNA Apak expression plasmid was co-transfected with lentivirus coated plasmids pSPAX2 and pMD2.G.The supernatant was collected, filtered, and used to infect HCT116 cells.The positive clones were screened out by puromycin culture and Western blot was used to detect Apak knockout cell lines.Luciferase reporter gene assay, flow cytometry analysis and colony formation assay were used to examine p53 activity and apoptosis of Apak knockout cells, respectively.Results Apak knockout HCT116 cell lines were generated in which p53 activity and apoptosis were increased,but the colony formation was decreased.Conclusion The Apak stable knockout cell lines of HCT116 are successfully generated by CRISPR/Cas9 system for further functional study.
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Aim To investigate the effects of isoliquiri-tigenin(ISL)on C6 glioma cell proliferation and differ-entiation.Methods C6 glioma cells’viability and proliferation were respectively measured by SRB test. Colony formation of C6 glioma cells from different groups was assayed.After culturing the cells from each group,giemsa staining was used to observe cell mor-phology.RT-PCR was applied to detect mRNA expres-sion of GFAP.Western blot was applied to detect the expression of GFAP.Results ISL effectively inhibited the viability of C6 glioma cells when compared with the control group in a concentration-dependent manner (P<0.01).The morphological observation under light mi-croscope showed that:in the control group,most of the undifferentiated C6 cells showed long fusiform and po-lygonal shape.Compared to the control group,the C6 cells treated with ISL revealed alteration in morphology such as astrocytes with smaller smooth,round body and much finer longer,tapering processes.The cloning for-mation rate detection revealed that:the colonies in the control group semerged earlier and were larger than those experimental ones,the cloning formation rate was higher,while almost no effective cells colony emerged in ISL treated groups(P <0.01 ).Western blot and RT-PCR analysis showed that GFAP expression in the ex-perimental groups increased(P <0.01).Conclusion ISL may inhibit the proliferation of C6 glioma cells and induce their differentiation.
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Objective To establish radiation?resistant lung carcinoma cell lines, and to investigate the changes in morphology, apoptosis, invasive migration, and epithelial?mesenchymal transition ( EMT) in cells. Methods The radiation?resistant lung carcinoma cell lines were obtained by exposure of lung carcinoma cell lines, A549 and H1299, to radiation with a low dose in fractions, a sublethal dose, or a gradually increasing dose. The morphological changes in cells, radiosensitivity, survival rates after exposure, apoptosis rates, changes in invasive migration, and expression of EMT marker proteins were evaluated using microscopy, colony formation assay, CCK?8 assay, flow cytometry, transwell migration assay, and Western blot, respectively. Results Radiation with a gradually increasing dose successfully induced the radiation?resistant cell lines, A549R and H1299R. The morphological study showed that the morphology of radiation?resistant cells was converted to the morphology of mesenchymal cells. Compared with A549 and H1299 cells, the values of D0 , Dq , and SF2 were significantly increased in A549R ( P=0.017,P=0.001,P=0.000) and H1299R (P=0.033,P=0.000,P=0.008) cells, respectively;the values of α and α/β were significantly reduced in A549R (P=0.018;P=0.007) and H1299R (P=0.001;P=0.009) cells, respectively. The survival rates in A549R and H1299R cells after exposure to radiation with various doses were significantly higher than those in the control groups (all P<0.05). After exposure, the apoptosis rates were significantly reduced in A549R and H1299R cells ( P=0.02,P=0.01);the invasion and migration rates were significantly increased in A549R (P=0.000;P=0.001) and H1299R (P=0.001,P=0.002) cells;the expression of E?cadherin was significantly down?regulated in A549R and H1299R cells (P=0.00,P=0.01), while the expression of vimentin was significantly elevated in A549R and H1299R cells ( P= 0. 02, P= 0. 01 ) . Conclusions The radiation?resistant lung carcinoma cell lines are successfully established. Both cell lines show enhanced invasion and migration, which may be associated with EMT.
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Social theory has provided a useful framework for research with microorganisms. Here I describe the advantages and possible risks of using a well-known model organism, the unicellular yeast Saccharomyces cerevisiae, for sociobiological research. I discuss the problems connected with clear classification of yeast behaviour based on the fitnessbased Hamilton paradigm. Relevant traits include different types of communities, production of flocculins, invertase and toxins, and the presence of apoptosis.
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Objective:To study the role of FDCs-miR-548m-CDK6 axis on clonogenicity in mantle cell lymphoma. Methods:RT-qPCR and Western blot were used respectively to test the expression of miR-548m and CDK6. Bioinformatics assay was applied to predict the targets of miR-548m, and Western Blot was used to test the expression level of CDK6 after miR-548m overexpression or in-hibition. Luciferase report assay was performed to test whether CDK6 was a direct target of miR-548m. Colony forming assay was used to test the colony forming activity in MCL after overexpression of miR-548m or knockdown of CDK6. Results:Cell adhesion to FDCs induced downregulation of miR-548m and CDK6 expression in MCL. Bioinformatics assay revealed that miR-548m could target the 3'-UTR of CDK6 and that a negative correlation exists between the level of miR-548m and the CDK6 expression. Luciferase report as-say confirmed that miR-548m directly targeted 3'-UTR of CDK6. Colony forming assay showed that overexpression of miR-548m or knockdown of CDK6 significantly suppressed MCL colony formation. Conclusion:This study reveals that FDC-enhanced mantle cell lymphoma clonogenicity is mediated by the miR-548m/CDK6 axis.
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Objective:This study aims to investigate the expression pattern of miR-512-3p in breast cancer and noncancerous paired specimens as well as the effects of miR-512-3p on the proliferation, apoptosis, cell cycle, and cloning of MD-MBA-231 breast cancer cells. The study also aims to identify the miR-512-3p target gene. Methods:Reverse transcription-polymerase chain reaction (RT-PCR) was conducted to quantify the miR-512-3p expression in breast cancer and noncancerous paired specimens. Methylthiazol tetrazolium (MTT) assay, flow cytometry, and clone formation assay were used to characterize the function of miR-512-3p in breast cancer. Target prediction was performed using the TargetScan, PicTar, and miRanda software. The results were validated using RT-PCR and western blot target validation. Results:The relative expression of miR-512-3p in breast cancer specimens was significantly lower than those in normal breast specimens (P<0.05). MTT assay revealed that 48 h after transfection, miR-512-3p significantly repressed the proliferation of MD-MBA-231 cells at a suppression rate of 45.38%and at a concentration of 100 nmol/L. MiR-512-3p increased the percentage of early apoptotic cells in the treatment groups (9.32 ± 0.41)%compared with those in the blank controls (3.1 ± 0.54)%and in the negative controls (2.9 ± 0.39)%(P<0.05). Significant differences were found in the percentages of the G0/G1-and G2/M-phase cells after miR-512-3p transfection compared with those in the controls (P<0.05). In the cloning assay, clone formation was inhibited in the miR-512-3p-transfected groups compared with those in the control groups. RT-PCR and western blot results indicate that miR-512-3p significantly inhibited the c-FLIP mRNA and protein expression. Conclusion:MiR-512-3p expression is relatively decreased in breast cancer specimens compared with those in the normal samples. The negative effect of miR-512-3p on cell proliferation and clone formation and its positive effect on early apoptosis through c-FLIP targeting suggest that miR-512-3p acts as a tumor suppressor gene in breast cancer. Therefore, miR-512-3p may be a new target for the diagnosis and treatment of breast cancer in the future.
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Objective To investigate the effects of resveratrol on cell growth,migration,and invasion of human breast cancer cell line MDAMB-231.Methods MTT assay and soft agar colony formation assay were used to detect the effects of resveratrol on cell growth.The effects of resveratrol on cell migration and invasion were determined by Transwell migration assay and Transwell invasion assay.respectively.Results Resveratrol(25 to 200 μmol/L)inhibited the growth of MDA-MB-231 cells in a time-and concentration-dependent manner(P＜0.01).The colony formation rate of MDA-MB-231 cells decreased with the increase in the concentrations of resveratrol.After being treated with resveratrol of 25,50,100 and 200 μmol/L for 24 hours,the invasive and migratory ability of MDA-MB-231 cells was significantly inhibited (P＜0.01).Conclusion Resveratrol could inhibit the growth of MDA-MB-231 cells,and the invasive and migratory ability of MDA-MB231 cells is also inhibited by resveratrol through degrading extracellular matrix.
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Objective:To observe the biological behaviors of colorectal cancer LS174 cells before and after pcDNA3.0-hugl-1 transfection,so as to investigate the association of hugl-1 with colorectal cancer.Methods:The eukaryotic expression vector pcDNA3.0-hugl-1 was constructed and transfected into LS174 cells.RT-PCR and Western blotting methods were used to analyze the expression of hugl-1 mRNA and protein in LS174 cells before and after transfection.Soft agar colony formation assay,wound-healing experiment,adhesion assay and Matrigel invasion assays were used to study the effects of hugl-1 expression on the proliferation,adhesion,movement and invasion in LS174 cells.Results:The recombinant plasmid pcDNA3.0-hugl-1 was successfully constructed.RT-PCR and Western blotting showed that the hugl-1 expression was higher in cells transfected with pcDNA3.0-hugl-1 than in those un-transfected or empty vector-transfected cells (P
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PURPOSE: Umbilical cord blood transplantation is a alternative method as new hematopoietic stem cell transplantation and has been performed clinically in indicated disease. However, it have the problems for long-term storage of cord blood in liquid nitrogen and for limited application to adult due to small amount of hematopoietic stem cell. Therefore, several centers have carried out active research for ex vivo expansion of cord blood stem cell. We investigated the hematopoietic function of cord blood plasma for development of new techniques. METHODS: We acquired the nucleated cells of cord blood from healthy infant and bone marrow from healthy donor received granulocyte-colony stimulating factor. We evaluated hematopoietic colony formation according to source of stem cell and plasma by semisolid culture medium. Three experimental groups were divided as source of plasma: group for cord plasma, group for bone marrow plasma, group for mixture of cord plasma and bone marrow plasma. RESULTS: The results were as follows: 1) The colony formation according to source of stem cell in commercialized standard semisolid culture medium showed that cord blood in the number of CFU-GM was less than bone marrow, but not significantly different in CFU-GEMM. 2) The colony formation according to source of stem cell in semisolid culture medium using experimental plasma showed that cord blood in the number of CFU-GM was more than bone marrow. There were no cytotoxic effect of plasma to experimental cells. 3) The colony formation in semisolid culture medium contained plasma according to experimental group showed that the number of CFU-GM in cord blood plasma was significantly more than bone marrow plasma in spite of different source of stem cell. Conclusions: These results suggested that cord blood might contain enough hematopoiesis to enable to perform transplantation compared with bone marrow and, also, cord blood plasma might be contributed more effective colony formation than bone marrow plasma. Therefore, we propose that it may be good to store cord blood cells with cord blood plasma in long-term storage. We will investigate the composition of hematopoietic growth factors and cytokines in cord blood plasma and the effect of cord blood plasma for ex vivo expansion of cord blood cells.