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@#Objective: To investigate the role of LncRNA RP11-513G11.1 in the chemoresistance and evaluation of prognosis in small cell lung cancer (SCLC). Methods: From June 2012 to June 2017，98 cases of SCLC tissue, 30 cases of paracancerous tissue and 30 cases of normal lung tissue were performed by surgery, puncture biopsy or bronchoscopic biopsy from the Affiliated Hospital of Southwest Medical University. QRT-PCR was used to detect the expression of LncRNA RP11-513G11.1 in SCLC tissue, paracancerous tissue, normal lung tissue and SCLC sensitive cell strain H69, drug resistance cell strain H69AR.All patients received EP regimen (etoposide+cisplatin). According to their chemosensitivity, they were divided into chemosensitivity group and drug resistance group. The expression of LncRNA RP11-513G11.1 in two groups was detected. The relationship between RP11-513G11.1 expression and prognosis, survival time and risk factors of OS in patients were analyzed. Results: The expression of LncRNA RP11-513G11.1 in H69AR chemoresistant cells (13.790±2.830) was significantly higher than that in H69AR chemosensitive cells (1.080±0.090) (P<0.01),the expression level of LncRNA RP11-513G11.1 in SCLC tissues (8.558±1.130) was significantly higher than that in adjacent tissues (1.188±0.090) and normal lung tissues (1.636±0.150) (all P<0.01), the expression of RP11-513G11.1 in chemoresistant patients was significantly higher than that in chemosensitive patients (4.974±0.313) (P<0.01). The expression of RP11-513G11.1 was not related to gender and age, but was related to disease stage, lymph node metastasis, distant metastasis and chemotherapy resistance in SCLC patients (all P<0.05); High expression RP11-513G11.1 patients was shorter PFS [(12.59 ±2.08) months] and OS [(24.98 ±1.56) months] than those with low expression [(25.47±1.23) months] and [(39.03±2.67) months] (P<0.01). Univariate and multivariate analysis showed that RP11513G11.1 expression, disease stage and distant metastasis were independent prognostic risk factors for SCLC patients (all P<0.05). Conclusion: LncRNARP11-513G11.1 may be a potential biomarker of chemosensitivity and prognosis in SCLC patients.
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OBJECTIVE:To study the effects of butin in Vernohia anthelmintica(VW)on proliferation of human immortal ke-ratinocyte cell strain HaCaT and cell secretory factors,and explore the mechanism of butin in VW in the treatment of vitiligo. METHODS:MTT method was used to determine the survival rate of HaCaT cells cultured by 0 (blank control),0.1,0.5,1.0, 5.0,10.0 μg/mL of butin for 48 h. Enzyme-linked immunosorbent assay was used to determine the contents of cell secretory factors as endothelin 1 (ET-1),ET-3,melanocyte stimulating hormone (MSH),stem cell factor (SCF),basic fibroblast growth factor (bFGF)in culture medium after HaCaT cells were cultured by 0.5,1.0,5.0 μg/mL of butin for 48 h. RESULTS:Compared with blank control,cell survival rate was increased to varying degrees after cultured by 0.1-5.0μg/mL of butin for 48 h,while decreased after cultured by 10.0 μg/mL of butin. Contents of ET-1,SCF,bFGF in culture medium were significantly increased after cultured by 0.5,1.0,5.0μg/mL of butin for 48 h(P<0.01);and contents of ET-3,MSH in culture medium were significantly increased af-ter cultured by 1.0,5.0 μg/mL of butin for 48 h(P<0.01). CONCLUSIONS:Butin can promote the proliferation of HaCaT cells, the mechanism may be associated with promoting the secretion of cell secretory factors of ET-1,ET-3,MSH,SCF,bFGF.
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OBJECTIVE:To study the effects of butin in Vernohia anthelmintica(VW)on proliferation of human immortal ke-ratinocyte cell strain HaCaT and cell secretory factors,and explore the mechanism of butin in VW in the treatment of vitiligo. METHODS:MTT method was used to determine the survival rate of HaCaT cells cultured by 0 (blank control),0.1,0.5,1.0, 5.0,10.0 μg/mL of butin for 48 h. Enzyme-linked immunosorbent assay was used to determine the contents of cell secretory factors as endothelin 1 (ET-1),ET-3,melanocyte stimulating hormone (MSH),stem cell factor (SCF),basic fibroblast growth factor (bFGF)in culture medium after HaCaT cells were cultured by 0.5,1.0,5.0 μg/mL of butin for 48 h. RESULTS:Compared with blank control,cell survival rate was increased to varying degrees after cultured by 0.1-5.0μg/mL of butin for 48 h,while decreased after cultured by 10.0 μg/mL of butin. Contents of ET-1,SCF,bFGF in culture medium were significantly increased after cultured by 0.5,1.0,5.0μg/mL of butin for 48 h(P<0.01);and contents of ET-3,MSH in culture medium were significantly increased af-ter cultured by 1.0,5.0 μg/mL of butin for 48 h(P<0.01). CONCLUSIONS:Butin can promote the proliferation of HaCaT cells, the mechanism may be associated with promoting the secretion of cell secretory factors of ET-1,ET-3,MSH,SCF,bFGF.
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Objective To explore the inhibitory effect of Oldenlandia diffusa extract on colorectal cancer angiogenesis in BALB/c mice. Methods Thirty-two BALB/c mice with subcutaneous CT26 colon cancer animal model were randomly equally divided into four groups,including the control group (groupⅠ,saline 0.1 mL/(10. d), O. diffusa ethanol extract of 90 mg/(kg.d) (groupⅡ), O. diffusa ethanol extract of 180 mg/(kg.d) (groupⅢ) and O. diffusa ethanol extract of 360 mg/(kg.d) (group Ⅳ) . Each group of mice were treated with intragastric administration of law administration 12 days after vaccination, then stopped and continue fed to 32 days, and the mice were killed. Micro-vascular dense ( MVD) was observed and countered under the microscopy by immunohistory chemistry. Results The murine colon tumor volumes of GroupⅡ,ⅢandⅣwere significantly less than that of groupⅠ,with significant difference ( <0.05) . The tumor microvessel density values of four groups was (7.83±2.87), (5.32±1.27), (1.77±0.70) and (1.87±0.68),respectively. The number of tumor blood vessels in GroupⅡ,Ⅲ and Ⅳ were significantly less than that of Ⅰ group, with significant difference ( <0.05) . Conclusion Within a certain dose range, the ethanol extract of O. diffusa can significantly inhibit the mouse colon cancer and the mechanism may be realated to inhibiting tumor angiogenesis.
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Objective To explore the inhibitory effect of the ethanol extract of Oldenlandia diffusa on the proliferation of CT-26 colon cancer cells which come from BALB/c mice. Method We determined the inhibitory effect of different concentrations of ethanol extract of Oldenlandia diffusa on CT-26 cells' proliferation by using methyl thiazolyl tetrazolium (MTT), and calculated the 50% inhibiting concentration (IC50) . Results As to the same concentration, the inhibitory effect of the ethanol extract of Oldenlandia diffusa on CT-26 cells was increased with time, for exsample:after treated with 0.08 mg/mL of ethanol extract of Oldenlandia diffusa for 24 h, 48 h and 72 h, the inhibitory rates of CT-26 cells were (16.67 ±9.35)%, (34.66 ±9.23)%and (40.07 ±9.16)%, respectively. After treating CT-26 cancer cells for 24 h, 48 h and 72 h, the IC50 values of the ethanol extract of Oldenlandia diffusa were 0.315,0.155 and 0.115 mg/mL, respectively. In the same treatment time, the inhibitory effect of the ethanol extract of Oldenlandia diffusa on CT-26 cells was increased with the increase of concentration:after treatment for 72 h with different concentrations (0.06 mg/mL,0.08 mg/mL,0.10 mg/mL,0.12 mg/mL, 0.14 mg/mL,0.16 mg/mL,0.18 mg/mL and 0.20 mg/mL) of the ethanol extract of Oldenlandia diffusa,the inhibitory rates of CT-26 cells were (35.46 ±3.59)%, (40.07 ±9.16)%, (40.77 ±6.92)%, (52.81 ±1.87)%, (54.22±2.35)%, (68.72±3.71)%, (70.04±8.03)%and (71.84±3.12)%, respectively. Conclusion The ethanol extract of olenlandia diffusa can inhibit the proliferation of CT-26 colon cancer cells from BALB/c mice in a time and dose dependent manner.
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Objective To evaluate the ability of 3-AB to sensitize the human esophageal carcinoma cell strain (CaEs-17) to radiation in v/tro and its mechanisms. Methods CaEs-17 cells were treated with 3-AB at 0, 2.5, 7.5 mmol/L and given irradiation O, 3, 6, 9, 12 Gy. 3-AB concentration in each group was made dose-survival curve using multi-target single-hit maiths model by clonogenie assay. MTT assay was performed to observe the survival of irradiated cells.comet assay and metaphase chromosome analysis were used to measure the DNA damage degree and chromosome aberration of CaEs-17 cell after 3-AB treatment and irradiation. Results Cell survival experiments showed SER of 1.21, 1.52 for 2.5 mmol/L, 7.5 mmol/L 3-AB respectively using multi-target single-hit maths model. The survival fraction of irradiated CaEs-17 cell was decreased after 3-AB treatment. DNA damage and the chromatid breakage number of irradiated CaEs-17 cells were increased after 3-AB treatment. Conclusions 3-AB, a PARP inhibitor, can enhance the radiosensitivity of human esophageal carcinoma cell strain (CaEs-17). DNA damage repair inhibition by 3-AB might be one of the mechanisms.
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Objective To study the characteristics of Z-HL_(16)C cell strain mutated from the human embryonic lung fibroblasts(HL).Methods Morphology observation,chromosome cytogenetics,tumorigenesis in nude mice,and the sensitivity to virus were examined.Results As a transformed cell strain,the Z-HL_(16)C showed polygon shapes liking epithelium;the number of chromosome was sub-triploid or hyper-triploid,and the structures of 30%～40% chromosomes were abnormal.The incidence of its inoculation in nude mice was 100%,and the pathological studies proved its tumorigenesis.The cytopathic effects of many kinds of viruses were observed in Z-HL_(16)C cell strain.Conclusion The Z-HL_(16)C is one of the cancer cell strains transformed from HL cells.
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Objective To explain the effects of C-reactive protein(CRP) on lectin-like oxidized low density lipoprotein receptor-1 expression on THP-1 derived macrophages and the related signal transduction pathways.MethodsTHP-1 cells were differentiated into macrophages with the stimulation of PMA.THP-1 derived macrophages were incubated with CRP and co-incubated with inhibitors of NF-?B、AP-1 and MARK signal transduction pathways.The expression of LOX-1 antigen and mRNA was analyzed by ELISA and RT-PCR.Results CRP stimulated the expression of LOX-1 antigen and mRNA on macrophages in a dose-dependent manner.NF-?B inhibitor BAY11-7085 suppressed the inducible effects of CRP on LOX-1 expression.Conclusion CRP increased LOX-1 expression on THP-1 derived macrophages at transcription and post-transcription levels.The NF-?B signal transduction pathway may be involved in such process.
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Glucose-6-phosphate dehydrogenase (G6PD) derives from the expression of the house-keeping gene G6PD. Recent studies have indicated that G6PD is related to tumor genesis, growth, clinical phenotype, therapy, and prognosis. To elucidate the relationship between G6PD and cancer, three siRNA sequences and one negative control sequence were designed based on the 3' noncoding region of the human G6PD gene. Two complementary single-strand DNA (sense and antisense) were designed and synthesized based on siRNA sequences. The DNA fragments were annealed and ligated to the GFP expression vector pRNAT-U6.2/Lenti. One siRNA with higher interference efficiency than the other two was found after siRNA plasmid transfecting human skin A375 melanoma cells. After lentivirus particle packaging and virus production, the A375 cells were infected, and the single cell clone was acquired and cultured to establish the stable cell strain. Western blotting showed that the endogenous G6PD in the stable A375 cell strain was 0.257 ? 0.074, which was 11.17% of G6PD expression (2.301 ? 0.286) in wild type A375 cells. The final siRNA interference efficiency in this stable cell strain was 88.83%. The G6PD activity of A375-G6PD?驻 was 21.53% of A375-WT. Further study showed that A375-G6PD△ doubling generation time prolonged, and its proliferation was greatly inhibited and the cloning efficiency lowered 25%(P
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Objective To obtain a lasting and purified cell line of vascular smooth muscle cells (VSMCs) by primary culture, and to provide these test materials related research. Methods The primary and transfer culture was done by tissue - piece inoculationand trypsin digestion,respectively. The cells were frozen in liquid nitrogen,and cultured cells were identified by phase contrast microscopy and immunohistochemical SP kit. Results 85% of inoculated tissue pieces survived. More than 90% of VSMCs regrew in transfered culture. The frozen cells could recover their proliferation by several culture transfers. The VSMCs showed the typical "peak and valley" characteristics under microscopy. Immunohistochemical staining with antibody against SM - a - actin expression in these cells was positive. Conclusion The tissue - piece inoculation is a simple method for obtaining satisfactory VSMCs in short term.
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OBJECTIVE: To observe the neuroprotective effect of Baicalein on H2O2-induced PC12 injury.METHODS: The PC12 cells cultured in vitro were preincubated with BAI for 24 h before being duplicated into the irreversible oxidative damages model with 500 ?mol?L-1 H2O2,then the protective effect of BAI (BAI presence or withdrawal) on these damaged cells in vitro was observed. The cell activity was detected by MTT microcolorimetry, and the lactate dehydrogenase(LDH) level in supernatant was determined as well. RESULTS: After pretreatment with BAI for 24 h,the injury of PC12 cells was significantly lessened in the presence or withdrawal of drugs in modeling, especially in the presence of drugs. Microscopically, PC12 cells can be seen with fuzzy edge and increased size, and cell fusion occurred in the BAI-treated H2O2 injury model. Cell activity increased,LDH level in the medium decreased, which were significantly different as compared with the model group. CONCLUSION: BAI can significantly reduce the hypoxic-ischemic injury of PC12 cells. Its mechanism may be related to its ability in reducing the accumulation of free radicals induced by blockage of the respiratory chain, antioxidation and free radical scavenging.
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Objective: To detect the ability of medicine serum of Changantai and its major components to kill Co26lu colon cancer cells in vitro by means of serum pharmacology.Methods: There were 5 groups of rats.5-Fu 30mg/kg were injected into the abdominal cavity of the rats in 5-Fu blood serum group and abdominal aorta blood was drawn 1 hour after administration.The other four groups were intragastrically administrated with Changantai(dry paste),rhubarb(dry paste),radix actinidiae(dry paste) and NS 0.6ml/kg/d respectively and the blood was draw twice,that was,2ml orbital vein hemospasia 1 hour after administration and abdominal aorta hemospasia 1 hour after the last administration of three consecutive days with asceptic serum segregation.Ability of medicine serum to kill tumor cells was observed by means of MTT method.Results: medicine serum of 5-Fu,Changantai and its components with different concentration had different ability in vitro to kill Co26lu cells,which was concentration dependent.Conclusion: Changantai can inhibit cancer cells from growing and metastasis.
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To construct an immortalized rat astrocyte strain genetically modified by rat preprogalanin gene (IAST/GAL) and detect its galanin (GAL) expression and secretion, a cDNA fragment of rat GAL in plasmid of pBS KS(+)-GAL was inserted into eukaryotic expression vector pcDNA3.1(+) by DNA recombinant technology, then the restriction enzyme digestion and DNA sequencing were carried out to evaluate the recombinant. The pcDNA3.1 (+)-GAL and pcDNA3.1 (+) construct were transfected into immortalized rat astrocyte strain (IAST) by lipofectamine and the population of cells which stably integrated the construct was selected with 600 μg/mL G418. Individual clones were screened and expanded into clonal cell strains. Detection of Neo gene was used to validate the success of the transfection. Immunocytochemical staining, RT-PCR and radioimmunoassay were used to detect the expression and secretion level of GAL. The recombinant had been successfully constructed by restriction enzyme digestion and DNA sequencing. Detection of Neo gene showed that the pcDNA3.1 (+)-GAL and pcDNA3.1 (+) have been successfully transfected into IAST. After selection by using G418, IAST/GAL and IAST/Neo cell strains were obtained.IAST/GAL, IAST/Neo and IAST were immunostained positively for GAL, but the GAL average optical density of IAST/GAL was significantly higher than that of IAST/Neo and IAST (P＜0.01). The level of GAL mRNA expression and the supernatant concentration of GAL in cultured IAST/GAL were significantly higher than those of IAST and IAST/Neo (P＜0.01), but no significant differences were found between the IAST and IAST/Neo (P＞0.05). It was concluded that IAST/GAL strain was constructed successfully and it might provide a basis for the further study of pain therapy.
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Objective To explore the relationship between drug resistance of leukemic cells and Caspase-3,this study took adriamycin(ADR)-resistant human chronic granulocytic leukemic cell strain K562/AO_2 as research subject,observing the cell survival and the morphological change of cell apoptosis under the action of ADR and arsenic sulfide and the Caspase-3 activity before and after putting in the Caspase-3 inhibitor.Methods ① The 3-(4,5-dimethylthiazo-2-yl)-2,5-diphenyl-tetrazolium bromide(MTT) method was used to determine the cell survival(A value) of K562/AO_(2)cell strain under the action of ADR and arsenic sulfide.② DNA agarose gel electrophoresis was performed to observe the DNA cleavage of apoptotic cells.③ The enzyme colorimetric activity assay(CAA) method was used to measure the change of the Caspase-3 activity of K562/AO_(2) cell strain.Results ① The A value of K562/AO_(2) cells had a time and dosage dependent relation with arsenic sulfide.② Apoptosis occurred in the K562/AO_(2) cell strain affected by arsenic sulfide.③ Compared with the cell strains with the Caspase-3 inhibitor added,the Caspase-3 activity of those without the Caspase-3 inhibitor increased remarkably(P
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Objective To discuss the growth suppressing, apoptosing effect of new type tumor-supressor gene-p33ING1 in stomach cancer cell strain, and to explore new strategies and methods in tumor therapy. Methods The PCDNA3/p33ING1 nuclear expressing microsome was constructed, p33ING1 and wt-p53 were implanted to human stomach cancer cell both and to evaluate the effect of p33ING1 and p53 toward stomach cancer cell and synergism between them. Results The PCDNA3/p33ING1 nuclear expressing microsome was successfully constructed. The human stomach cancer cell strain SSCG-7901 under implantation of p33ING1 and wt-p53 showed a significant decrease in cell growth, the coupling time was delayed, DNA synthetic phase was shortened and G0/G1 phase prolonged. The cell collapse increased. Conclusions Despite of the tumor-inhibiting effect and biochemical activation of p33ING1, it also plays a role with p53 gene in controling growth of stomach cancer cell, inducing cell collapse and hampering cell proliferation cycle. P33ING1 and p53 are synergistic to each other.
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This experiment was performed to elucidate the cytologic origin of chemically induced MFH in Wistar rats. The tumor was produced by injections of DMBA(9,10-dimethyl-1,2-benzanthracene). With the produced MFH, cell culture and cloning were performed, followed by establishment of a cell strain, which was investigated by immunohistochemical and electron microscopic studies. The results were as follows. A) By immunohistochemistry of the tumor tissue, fibroblastic cells were positive for MEP-1(specific antibody for fibroblastlike cell of MFH, Takeya, 1993) and Anti-hPH(beta)(Anti-prolyl 4-hydroxylase beta), but negative for TRPM-3 and F4/80. Histiocytelike cells were positive for TRPM-3 and F4/80, but negative for MEP-1 and Anti-hPH(beta). In immunoelectron microscopy, normal spleen macrophage showed linear reactivity in cell membrane for TRPM-3, whereas histiocytelike cells of the tumor disclosed negative reaction. B) At 5 weeks of the primary tumor cell culture, the cells exhibited typical storiform pattern of MFH. C) The established cell strain revealed immunoreactivity for MEP-1 and Anti-hPH(beta), but negative for TRPM-3. The cloned tumor cells showed morphologic characteristics of undifferentiated fibroblastic cell. Latex particle (0.80 micrometer size) phagocytosis was negative in the cloned cell strain. The results of the current study support the concept that principal component cells of MFH is of fibroblastic cell origin.
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Animales , Ratas , 9,10-Dimetil-1,2-benzantraceno , Técnicas de Cultivo de Célula , Membrana Celular , Células Clonales , Clonación de Organismos , Fibroblastos , Histiocitoma Fibroso Maligno , Inmunohistoquímica , Macrófagos , Microscopía Inmunoelectrónica , Microesferas , Fagocitosis , Ratas Wistar , BazoRESUMEN
Objective: To investigate the effect of recombinant BCG secreting IL-2 (rBCG-IL-2) on peritoneal macrophages of mouse and inquire into the effect of rBCG-IL-2 on immunity of host. Methods: Gene engineering technology, electricity transfer- ence and inoculation tumor cell are adopted; MTT assay is used to determine tumoricidal effect of macrophages. Results: Tumo- ricidal effect of macrophages of rBCG-IL-2 groups was higher than those of the control coups and BCG groups, and the tumorici- dal effect of macrophages could reach a high state in a week. Conclusion: rBCG-IL-2 can raise tumoricidal effect of macrophages, improve immunity of host and is a prospect biological preparation.
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AIM: To investigate the influence of matrine on cyclin E and cyclin A in U937 cell strain. METHODS: The inhibitory effect of matrine on proliferation in U937 cell strain was observed by MTT.The distribution of cell cycle was measured by flow cytometry.Immunocytochemical method and Western blot were used to detect the protein expression of cyclin E and cyclin A. RESULTS: After being treated with matrine,the proliferation of U937 cell strain was inhibited significantly,the inhibiting effect conformed to time and dose-dependence,cells were arrested in S-phase.The expression of cyclin E and cyclin A was down-regulated significantly in a dose-dependence manner. CONCLUSION: Matrine could reduce the expression of cyclin E and cyclin A and inhibit the proliferation in U937 cell strain.
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OBJECTIVE:To study the inhibitory effects of propolis on growth of transplantation tumor in mice.METHO_ DS:Using different concentrations of propolis to feed the mice for two months,the tumor cells(S 180 )were transplanted into subaxillary tissue of the mice.After8days,the tumor mass was takent off the body of the mice,and weighted,then paraffin sections were observed and the number of karyokinesis of tumor cells was counted under the microscops.RESULTS:The weight of tumor mass were lighter in the propolis group than in the control group(P
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Aim To observe the cytotoxic and proliferous effects of Nitraria tangutorum flavone(NTF)on human A-704 strain cells.Methods In vitro cultured human A-704 strain cells,the cytotoxic and proliferous effects of NTF on human A-704 strain cells were determined with methods of trypan-blue and clone formation,and the microelement in cell was detected with X-electron microscope scanning.Results ① The trypan-blue detection showed that the living cells in the NTF group(0.1,0.5 and 5 mg?L-1)and the 5-FU group were decreased by 33.8%~57.3%,35.8%~80.1%,60.8%~99% and 40.3%~59.6% at 6~18 hour after treatment,which significantly differed from those of the control group(P