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Objective:To analyze and evaluate the application of quantitative fecal immunochemical test(FIT) in opportunistic screening of colorectal cancer in asymptomatic population undergoing health checkups.Methods:From January 1, 2018 to December 31, 2021, at the Health Management Center of the First Affiliated Hospital of Soochow University, 53 319 subjects who underwent routine health checkups and with quantitative FIT opportunistic screening for colorectal cancer were selected. Those with positive quantitative FIT results and received colonoscopy were enrolled in the FIT positive group, and those with negative quantitative FIT results and received colonoscopy were enrolled in the FIT negative group. The participation rate and positive rate of quantitative FIT were analyzed. The results of colonoscopy and pathological findings were taken as the gold standard, including normal, non-polyposis lesions, polyposis (hyperplastic and(or) inflammatory polyps, non-advanced adenoma, advanced adenoma), and colorectal cancer, the detection rates of various lesions of the FIT positive and negative groups, the quantitative FIT measurement value of subjects, and the sensitivity and negative predictive value of quantitative FIT for colorectal cancer and advanced adenoma were analyzed. The receiver operating characteristic curve (ROC) was drawn and the area under the curve (AUC) was calculated, the screening efficacy of quantitative FIT for colorectal cancer and advanced adenoma was evaluated. Chi-square test or Fisher exact probability method and Wilcoxon rank sum test were used for statistical analysis.Results:A total of 51 420 cases had completed quantitative FIT, and the total participation rate was 96.44% (51 420/53 319). Quantitative FIT was positive in 2 483 cases (4.83%). The participation rate of colonoscopy in FIT positive group was 26.22% (651/2 483), of which 540 cases were enrolled in FIT positive group. The colonoscopy participation rate of FIT negative group was 1.18% (576/48 937), of which 523 cases were enrolled in the FIT negative group. The detection rates of colorectal cancer and advanced adenoma in FIT positive group were both higher than those of the FIT negative group(3.9%, 21/540 vs. 0, 0/523; 16.1%, 87/540 vs. 3.3%, 17/523), and the differences were statistically significant(Fisher exact probability method and χ2=49.79; both P<0.001). Populations with quantitative FIT values from high to low were those with colorectal cancers, advanced adenomas, non-polyp lesions, non-advanced adenomas, normal, and hyperplastic and (or) inflammatory polyps (1 052.0 ng/mL(390.5 ng/mL, 3 058.0 ng/mL); 294.5 ng/mL (116.8 ng/mL, 951.8 ng/mL); 131.5 ng/mL (10.5 ng/mL, 327.3 ng/mL); 97.0 ng/mL (11.0 ng/mL, 238.0 ng/mL); 20.0 ng/mL (0.0 ng/mL, 175.3 ng/mL); 14.0 ng/mL (0.0 ng/mL, 171.0 ng/mL)), and the difference was statistically significant( H=120.53, P<0.001). The sensitivities(95% confidence interval (95% CI)) of quantitative FIT in colorectal cancer and advanced adenoma were 100.0%(80.8% to 100.0%) and 83.6%(74.8% to 89.9%), respectively. The negative predictive values (95% CI) were 100.0%(99.1% to 100.0%) and 96.7%(94.7% to 98.0%), respectively. The results of ROC analysis showed that the AUCs(95% CI) of quantitative FIT in colorectal cancer and advanced adenoma were 0.874(0.820 to 0.928) and 0.723(0.675 to 0.770), respectively. Conclusions:In this study, the participation rate of quantitative FIT is high. More patients with advanced adenomas and colorectal cancers are found in the high risk popolation with positive quantitative FIT. Quantitative FIT has a good sensitivity and a negative predictive value for colorectal cancer and advanced adenoma. Therefore, positive quantitative FIT-colonoscopies sequential screening should be advocated in population undergoing health checkups for colorectal cancer screening, and it may be applicable to large-scale population screening in China.
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Objective@#To evaluate the protective effect of granulocyte-colony stimulating factor(G-CSF) on neonatal rats after hypoxic-ischemic brain damage(HIBD)and its effect on endoplasmic reticulum (ER) stress.@*Methods@#According to the random number table, a total of 54 Sprague-Dawley (SD) rats aged 7 days were divided into 3 groups(18 rats in each group): Sham group, HIBD group and G-CSF group, and the improved Rice method was used to establish a neonatal rat model of HIBD.A dose of 50 μg/kg of G-CSF was administered intraperitoneally 1 hour after HIBD (G-CSF group), while the rats in HIBD group and Sham group received saline only.At 24 hours of HIBD, pups were euthanized to quantify brain infarct volume by using 2, 3, 5-Triphenyltetrazolium chloride.Hematoxylin-Eosin (HE) staining was used to observe the changes of brain structure.Neuronal cell death was determined by using terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL). Then the expressions of glucose-regulated protein 78 (GRP78), cysteinyl aspartate specific proteinase 12 (Caspase-12), CCAAT/enhancer binding-protein homologous protein (CHOP) were assessed by Western blot and immunofluorescence staining.@*Results@#Twenty-four hours after operation, HE staining showed that no significant neuronal damage was observed in Sham group.The brain tissue structure of rats in the HIBD group was significantly damaged, while some improvement was observed in the G-CSF group.The infarction volume in HIBD group[(25.40±5.15)%] increased compared with that in the Sham group[(0.31±0.15)%] and the G-CSF group[(16.36±4.97)%], and the differences were statistically significant(all P<0.05). There was increased positive expression of GRP78 protein in HIBD group, compared with that in the Sham group and the G-CSF group[(49.38±10.06)% vs.(9.12±4.50)%, (30.61±6.35)%], and the differences were statistically significant (all P<0.05). The percentage of apoptosis in the hippocampal CA1 region and conex in HIBD group [(44.84±11.54)%, (48.23±14.07)%] were higher than those in the G-CSF group [(17.87±7.20)%, (26.18±9.96)%], and the differences were statistically significant (all P<0.05). The expression of GRP78, CHOP and Caspase-12 in the HIBD group (0.63±0.24, 0.72±0.21, 0.68±0.25) were higher than those in the Sham group (0.20±0.08, 0.28±0.08, 0.23±0.07), and the G-CSF group (0.39±0.13, 0.51±0.18, 0.48±0.16), and the differences were statistically significant (all P<0.05).@*Conclusions@#G-CSF exerts neuroprotective effect on the neonatal rats after HIBD.G-CSF may be an effective treatment of HIBD by reducing ER stress-induced neuronal apoptosis.
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Objective@#To investigate the role of granulocyte-colony stimulating factor (G-CSF) on the regulation of inflammatory cytokines in neonatal hypoxic-ischemic brain damage(HIBD) rat model, and to explore the possible mechanism involved in G-CSF neuroprotective effect via the mammalian target of Rapamycin/p70 ribosomal S6 protein kinase (mTOR/p70S6K) signaling pathway.@*Methods@#A group of postnatal day 7 (P7) Sprague-Dawley rat pups (90 cases) were randomly divided into sham-operated group, hypoxia-ischemia(HI) group, G-CSF group, Rapamycin (RAP) group and control group, and the improved Rice method was used to establish a neonatal rat model of HIBD.One hour before HI induction, Rapamycin was administered intraperitoneally with a dose of 250 μg/kg, and the control group was given equal volume of ethanol injected intraperitoneally.One hour after HI, a dose of 50 μg/kg of G-CSF was injected intraperitoneally into the G-CSF group, Rapamycin group and control group.The same volume of normal saline was injected intraperitoneally into HI group and sham-operated group.Forty-eight hours after HI, Western blot was used to detect the protein levels of tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, IL-10, and the mTOR/p70S6K signaling pathway in brain tissue.Neuron injury of the hippocampal CA1 region and the cortex was assessed by Nissl staining, and infarct volume detected by 2, 3, 5-triphenyltetrazolium chloride staining.@*Results@#The G-CSF group and control group were associated with significantly reduced infarction volume compared to the HI group [(12.87±1.54)%, (11.90±1.31)% vs.(24.21±3.28)%], and the differences were statistically significant(P<0.05). There was an increased positive neuron cell number in the ipsilateral hemispheres of the hippocampal CA1 region in the G-CSF group and the control group [(61.00±4.90) cell/field and (61.67±6.40) cell/field] and cortex [(92.67±6.68) cell/field and (90.17±4.45) cell/field] compared with those in HI group [(42.62±4.46) cell/field and (70.83±6.97) cell/field], and the differences were all statistically significant (all P<0.05). The expression levels of TNF-α and IL-1β were significantly decreased in the G-CSF group and the control group, compared with those in HI group(TNF-α: 0.67±0.07, 0.55±0.05 vs.0.86±0.05; IL-1β: 0.65±0.06, 0.52±0.10 vs.0.86±0.06), and the differences were all statistically significant (all P<0.05). There was increased expression levels of IL-10, p-mTOR/mTOR and p-p70S6K/p70S6K in the G-CSF group and the control group, compared with those in HI group (IL-10: 0.68±0.04, 0.62±0.05 vs.0.34±0.02; p-mTOR/mTOR: 0.53±0.02, 0.51±0.01 vs.0.26±0.01; p-p70S6K/p70S6K: 0.89±0.03, 0.90±0.03 vs.0.55±0.02), and the differences were all statistically significant(all P<0.05). There was an increased infarct volume in Rapamycin group [(25.70±1.50)%], compared with the G-CSF group and the control group, and there were decreased number of positive neuron cell count in the hippocampal CA1 region [(40.67±3.50) cell/field] and cortex [(68.33±8.17) cell/field], increased expression levels of TNF-α and IL-1β (0.97±0.06 and 0.98±0.10, respectively), decreased expression levels of IL-10, p-mTOR/mTOR and p-p70S6K/p70S6K (0.21±0.02, 0.30±0.01 and 0.55±0.01, respectively) in the Rapamycin group, and the differences were all statistically significant (all P<0.05).@*Conclusions@#G-CSF may inhibit inflammatory responses after HIBD by up-regulating the mTOR/p70S6K signaling pathway in neonatal HI encephalopathy.
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Thiamin pyrophosphate (TPP) is a thiamine (vitamin B1) derivative and an essential cofactor in oxidative metabolism of the sugars,fatty acids and amino acids in living cells.By now,numerous TPP-dependent artificial riboswitch systems have been developed to regulate target gene expression but limited in bacteria,fungi or plant cells.Herein,the activating (switch-on) and inhibiting (switch-off) TPP-depended hammerhead ribozyme switches,which are from previous reported structures of prokaryotes screening,were investigated in mammalian cells.These ribozyme switches were inserted into the 3'UTR of the enhanced green fluorescence protein (EGFP) gene to construct the efficient ribozyme-based artificial switches through overlap extension PCR cloning.The HEK293 cells were transfected with the engineered ribozyme switches at increasing concentration of TPP.The EGFP gene-regulatory ability was analyzed with fluorescent microscope and flow cytometry.These TPP-inducible gene regulation devices showed the obvious ligand dose-dependency and excellent specificity.Two switch-on and one switch-off constructs demonstrated 3.1-fold or 1.9-fold increment and 2.3-fold reduction of EGFP level respectively with 150 μ mol/L TPP.The ligand-responsive ribozyme switches,by tuning the change of TPP concentration into the visual reporter genetic expression in cells,enable an efficient development of label-free,noninvasive and high-specific biosensors in living mammalian cells.
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This article reviews the cause of trastuzumab in treating human epidermal growth factor re-ceptor-2 (HER2) positive breast cancer, the evidence-based practice of continued use of trastuzumab, and some approaches to improve the efficacy of trastuzumab.
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Objective To study the effects of midkine(MK)gene small interfering RNA(siRNA)on adhesion and invasion of human breast cancer cells.Methods Real time PCR was used to evaluate MK mRNA expression in 7 human breast cancer cell lines Bcap-37,LCCI,MCF-7,MDA-MB-231,MDA-MB-435,MDA-MB-468,and ZR75-1.The cell line in which MK expression was the highest was transfected with different doses of MK siRNA.The expression of MK mRNA and protein was determined by real-time quantitative PCR and immunoflurescence staining.The cell adhesion was evaluated by MTT assay and invasion was examined by Boyden chamber method.Results Cell line MCF-7 expressed the highestlevel of MK mRNA in the 7 tested breast cancer cell lines.After being transfected with MK siRNA,MK mRNA and protein level of MCF-7 decreased in timeand dose-dependent manners.The adhesive and invasive ability of MCF-7 cell transfected with MK siRNA decreased in a dose dependent manner(P<0.01,P<0.01).Conclusions MK gene might play an important role in adhesion and invasion of human breast cancer cells.siRNA transfection could effectively inhibit adhesion,migration,and invasion of human breast cancer cell.
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Objective To study effects of polo-like kinase-1(PLK1)small interfering RNA(siRNA)on proliferation and apoptosis of undifferentiated human thyroid cancer cells.Methods 5 PLK1 siRNA(S1,S2,S3,S4 and S5)were constructed and used to transfect human thyroid cancer cell line ARO.RT-PCR was employed to pick out the most effective siRNA,which was then used to transfect ARO cell.RT-PCR and western blot were used to detect PLK1 expression in thyroid cancer cells,which were divided into different groups.MTT assay was performed to examine the effects of PLK1 siRNA on thyroid cancer cells in all groups.Apoptosis of thyroid cancer cells was observed by caspase-3 activity and TUNEL.Results All the 5 siRNA down-regulated PLK1 mRNA expression.among which S4 showed the best effect.S4 transfection could obviously inhibit proliferation of thyroid cancer cells in dose and time dependent manner.Compared with control groups,caspase-3 activity of cancer cells in s4 transfeeted group increased significantly.The effect of S4 transfection was dose and time dependent.TUNEL results showed apoptosis of cancer cells transfected by S4 siRNA was obvious and apoptosis of cells was dose-dependent.Conclusions PLK1 may play an important role in proliferation of undifferentiated thyroid carcinoma.PLK1 siRNA transfection can inhibit proliferation of throid cancer cell through apoptosis induction.
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ObjectiveTo study the effects of tumor-associated calcium signal transducer-2 (TROP-2) gene small interfering RNA(siRNA) on adhesion and invasion of human breast cancer cell. MethodsReal time PCR was used to evaluate the TROP-2 mRNA of seven human breast cancer cell lines Bcap-37 ,LCC1 ,MCF-7 ,MDA-MB-231,MDA-MB-435, MDA-MB-468 ,and ZR75-1. The cell line of TROP-2 highest expression was transfected with different dose of TROP-2 siRNA. The expression of TROP-2 mRNA and protein were determined by Real-time quantitative PCR and immumoflurescence method. The cell adhesion was evaluated by MTT assay,and invasion was exmined by hoyden chamber,respectively. Results Cell line MCF-7 showed the highest elevation of TROP-2 mRNA in seven breast cancer cell lines. The results from real-time quantitative PCR and immumoflurescence method showed that TROP-2 mRNA and protein reduced in time-and dose-dependent manners( P ＜ 0.01 ;P ＜ 0.01 ). The adhesive rate of siRNA groups(5 nM,10 nM,and 20 nM)was(52.9 +2.5)％ ,(25.6 ±2.3)％, ( 12.8 +2.2)％ (P ＜0.01 ) ,respectively.The transwell results showed that the invasion cells was(78 ± 17), (39 ± 15), ( 19 ± 16), ( 136 +25 ) and( 139 ±21 )in different groups(5,10,20 nM siRNA,and controls) ,respectively(P ＜0.01). ConclusionTROP-2 gene might play an important role in adhesion and invasion of human breast cancer cell. siRNA targeted TROP-2 could effectively inhibit adhesion and invasion of human breast cancer cell.
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ObjectiveTo study the scope of excison in breast-conserving surgery for breast carcinoma.MethodsClinical data of 275 breast cancer patients undergoing breast-conserving surgery in t he First Affiliated Hospital of Nanjing Medical University,the Affiliated Zhenjiang Hospital of Jiangsu University and Changzhou Traditional ChineseMedicine Hospital were retrospectively analyzed.The operation procedure and postoperative adjuvant therapy were carried out with the same protocol.Local and general conditions of patients were followed up regularly.Results271 out of 275 patients got follow-up.The follow-up rate was 98.5％.The follow-up time ranged from 1 month to 117 months,median follow-up time was 34 months.Six patients died of distant metastasis,2 with local recurrence.The 1-year,3-year,and 5-year overall survival rates were 99.5％,98.1％,and 95.7％,respectively.ConclusionsIt is safe to excise 1 cm normal breast tissue with clear margin confirmed by frozen section,followed by postoperative adjuvant therapy,endocrine therapy,and radiotherapy,this improves the life quality of patients with breast cancer.It is safe and effective to determine whether the disease is multicentric or multifocal by mammogram plus clinical breast examination.
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Objective To study the impact of TDGF-1 gene silience by small interfering RNA(siRNA)on the invasion and migration of human breast cancer cell. Methods 3 siRNA fragments were designed according to the characteristic of TDGF-1 gene sequence and the most appropriate siRNA was selected by fluorescence real-time quantitative RT-PCR method. After the human breast cancer cell line MDA-MB-468 was transfected by the selected TDGF-1 siRNA, mRNA and protein of TDGF-1 were determined by real time quantitative RT-PCR and western blot respectively. The migration and invasion ability of the cancer cell were evaluated by wound-healing assay and Boyden chamber model respectively. Results siRNA could down-regulate the level of mRNA and protein of TDGF-1 in a dose-and time-dependent manner. In vitro experiment showed that TDGF-1 siRNA transfection can effectively inhibit the clonal growth, invasion and migration of breast cancer cell in a dose-dependent manner. Conclusions TDGF-1 gene may play an important role in the migration and invasion of human breast cancer cells. siRNA transfection can inhibit the invasion of human breast cancer cells.
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Objective To evaluate our experience of minimal access parathyroidectomy for primary hy-perparathyroidism (PHPT) caused by parathyroid adenoma. Methods From Jan 2000 to Jan 2009, clinical data of 28 cases of PHPT treated by minimal access surgery were analyzed retrospectively. Results All patients except 3 had severe clinical manifestations, serum calcium and parathyroid hormonal (PTH) of all the cases were elevat-ed. The rate of accuracy of preoperation localization by the sestamibi scan combined with ultrasound was 100%.The pathological diagnoses of all the 28 cases were uni-gland parathyroid adenoma. Within the follow-up of 6 months, no case suffered complication and recurrence, the levels of serum calcium were normal or decreased slightly, and the levels of PTH were normal or elevated slightly but no higher than 2 times of normal PTH level. Conclusions Preoperative localization is very helpful by using combination of sestamibi and ultrasound scan. Minimally invasive parathyroidectomy is a kind of improving procedure for the localized parathyroid adenoma.
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Objective To discuss how to determine the operative margin in breast-conserving surgery in patient with early-stage breast cancer. Methods The breast-conserving surgery was performed on 111 patients with early-stage breast cancer who met the indication of breast-conserving. The operative incision was performed on normal breast tissue 1cm from the tumour edge and then the frozen sections of the surgical margin tissue were done to confirm that tumour was completely excised. The individualized therapy was performed according to pathologic examination and clinical stage. All patients were followed up regularly. Results 111 patients with early stage breast cancer received breast-conserving surgery and all had good curative effect except one patient had a local recurrence during the follow-up. Conclusions Operative incision on normal breast tissue 1cm from the tumour edge and frozen section of the surgical margin tissue are two crucial procedures to ensure the success of breast-conserving surgery and increase the life quality of patient with breast cancer.
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Objective To explore the safety of total thyroidectomy surgery on patients with thyroid carcinoma. Methods From Jan 1986 to Dee 2006, clinical data of patients who underwent total thyroidectomy (total thyroidectomy group) and sub-total or near-total thyroidectomy surgery (control group) for thyroid carcinoma were retrospectively analyzed to identify the incidences of complications, recurrent laryngeal nerve paralysis (RNLP) and secondary hypoparathyroidism in the two groups. Results In the control group there were 433 thyroid carcinoma patients who underwent sub-total or near-total thyroidectomy. Transient unilateral RLNP(13 eases), permanent unilateral RLNP(5 eases), transient hypoparathyroidism (11 cases) was diagnosed. There was no permanent hypoparathyroidism in this group. In the 70 cases of thyroid carcinoma patients receiving total thyroidectomy, there were 4 eases suffering from transient unilateral RLNP, one case from permanent unilateral RLNP (P > 0.05), and there were 7 eases from transient hopyparathyroidim (P < 0.01), 2 eases from permanent hypoparathyroidism (P < 0.05). Conclusion The incidence of RLNP after total thyroidectomy was not higher than that after subtotal or near-total thyroideetomy. Postoperative hypocalcaemia was the most common postoperative complication of total thyroidectomy. It is our belief that total thyroidectomy should be performed in selected patients.
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Objective In China primary hyperparathyroidism is not a kind of common disease as in the wesyrn countries.This article reports the current status in the diagnosis and treatment of primary hyperparathyroidism in the mainland of China. Methods We collected 730 cages of primary hyperparathyroidism diagnosed and treated in 7 top hospitals for endocrine surgery from 1965 to 2005.Results In this study.652(89.3%)cases were clinically symptomatic while 78(10.7%)cases were asymptomatic:442 cases were positive on 99mTc-MIBI scanning.Bilateral explorations were undertaken in 377 patients and unilateral or uni-gland exploration through the conventional incision in 204 cases.Minimally invasive parathyroidectomy in 143 cases.Endoscopically assisted 2 cm incision was taken in 6 cases for unilateral gland exploration.Pathologically 632(86.6%)cases were identified as adenoma,58(8.3%)cases were of hyperplasia and 40(5.5%)cases were of carcinoma.There were no major postoperative complications.While 20 patients suffering from recurrence or persistent postoperative hyperparathyroidism,the others are of normal or depressed serum level of calcium. Conclusions Preoperative localization is very helpful: Unilateral exploration for parathyroid adenoma is feasible; minimally invasive parathyroidectomy throush minimal incision is a kind of improving procedure for the localized parathyroid adenoma.
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Objective: To detect tumor cells in the peripheral blood of patients with breast cancer by usingthe MAGE-A1 and MAGE-A3 genes as specific tumor markers. Methods: Peripheral blood was obtained from 40 patients with breast cancer and 20 patients with benign diseases. The mRNA of the MAGE-A1 and MAGE-A3 genes in the peripheral blood mononuclear cells (PBMC) was detected by nested RT-PCR. The MAGE-A1 and MAGE-A3 transcripts in breast cancer were detected by RT-PCR. Results: Of the 40 breast cancer patients, MAGE-A1 and MAGE-A3 mRNA were positive in 12.5% (5/40)and 17.5%(7/40)of PBMC, respectively, and in 15.0 %(6/40)and 22.5 %(9/40)of breast cancer tissues, respectively. In the PBMC of the 40 breast cancer patients, 10(25.0%)samples were detected to express at least one type of MAGE mRNA. MAGE mRNA were not detected in the PBMC from the patients whose tumors did not express the MAGE genes, nor in the PBMC from the 20 patients with benign diseases. The positive rate of MAGE mRNA in the PBMC was closely correlated with the TNM stages. Conclusion: MAGE-A1 and MAGE-A3 mRNA could be specifically detected in the PBMCof breast cancer patients by our methods. They may be used as specific tumor markers for the detection of the circulating breast cancer cells.
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<p><b>OBJECTIVE</b>To study the major histocompatibility complex class II (MHC II) antigen and costimulatory molecules expression on the surface of breast cancer cells.</p><p><b>METHODS</b>MHC II antigen and costimulatory molecule expression on five breast cancer cell lines including MCF-7, SK-BR-3, T47D, MDA-MB-435 and ZR-75-30 were detected through flow cytometery analysis, with their expression level compared with that of normal mammary cell line HBL-100.</p><p><b>RESULTS</b>The MHC II expression level of the five breast cancer cell lines were significantly different from that of HBL-100 (P < 0.05). MHC II antigen expression of MCF-7 cells which was about 20 percent of HBL-100 was the lowest. MDA-MB-435 and ZR-75-30 cell expression levels were twice as much as that of HBL-100, with the fluorescence intensity of MDA-MB-435 the highest of all cells. CD40 molecule expression on the surface of MDA-MB-435 cells was the lowest, which was nearly ten percent of that of MCF-7 and HBL-100 cells. CD80 and CD86 molecule expression showed no difference in MDA-MB-435 or HBL-100 cell (P > 0.05), and those of the other four breast cancer cells were lower than that of HBL-100 (P < 0.05).</p><p><b>CONCLUSION</b>MHC II antigen and costimulatory molecule expression on the surface of breast cancer cells is abnormal, with different molecule expression in different cells. Breast cancer cells can escape immune surveillance through abnormal molecule expression.</p>
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Feminino , Humanos , Antígenos CD , Antígeno B7-1 , Antígeno B7-2 , Neoplasias da Mama , Alergia e Imunologia , Antígenos CD40 , Membrana Celular , Alergia e Imunologia , Antígenos de Histocompatibilidade Classe II , Glicoproteínas de Membrana , Células Tumorais CultivadasRESUMO
Background and purpose:The scarcity in quantity and continuous differentiation ability severely compromise the isolation and purification of cancer stem cells(CSCs).The existing sorting methods can only enrich CSCs as a heterogeneous population.However,more primitive CSCs closer to the source still have not been identified.Based on the CSC hypothesis,we hereby explored the possibility of isolation,culture,purification and identification of murine breast cancer stem cells in suspension culture combined with anticancer regimens. Methods :TM40D murine breast cancer cells were cultured in serum-free medium.The expression of CD44 + CD24-was measured by flow cytometry.Cells from the passage of TM40D cells with the highest expression of CD44 + CD24-were treated in combination with anticancer agents pacilitaxel and epirubicin at different peak plasma concentrations(PPC) for 24 hours,and then maintained under suspension culture.The rate of apoptosis was examined by flow cytometry with Annexin-V FITC/PI double staining method.Selected cells in different amounts were injected subcutaneously into BALB/C mice to observe tumor formation. Results :Cells of passage 10 in suspension culture had the highest percentage of CD44 + CD24-(about 77 percent).As few as 100 cells in 0.35 PPC could generate tumors in BALB/C mice. Conclusions :TM40D cell line contains breast cancer stem cells.This study suggested that suspension culture combined with anticancer regimens is a feasible approach for screening tumor stem cells.
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Objective To study the expression of Mage A1、A2、A3 in breast cancer tissues and four breast cancer cell lines Methods The expression of Mage A1、A2、A3 in breast cancer tissues and four breast cancer cell lines, MCF 7、Sk Br 3、MDA MB 435s and TM40D was detected by reverse transcription polymerase chain reaction (RT PCR) Results The positive expression rate of Mage A in breast cancer tissues was 13/33 (39%), of Mage A1 was 4/33 (12%), of Mage A2 was 8/33 (24%), and of Mage A3 was 7/33 (21%), respectively Both Mage A1 and Mage A3 were positive in breast cancer cell line MCF 7 and Sk Br 3 MDA MB 435s expressed Mage A2 and Mage A3 Mage A3 was positive in TM40D Conclusion Mage genes were often expressed in breast cancer, but expression of Mages varies in the breast cancer cell lines Mage genes encoding proteins are eligible for Mage peptide based active immunotherapy
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Objective To investigate the relationship between the expression of VCAM-1 and oncogenesis, tumor angiogenesis and metastasis in breast carcinoma. Methods Using semi-RT-PCR and immunohistochemistry technique, specimens from 63 patients of breast cancer,20 of fibroadenoma were detected for expression of VCAM-1.Microvessel density(MVD) was counted by endothelial cells immunostained using monoclonal antibody CD34.Circulating sVCAM-1 concentrations were measured by enzyme linked immunosorbent assay. Results Of 63 breast cancer tumor tissues VCAM-1 mRNA was detected in 37 cases. The rate of VCAM-1 mRNA in the primary breast cancer tissues were significantly higher than that in the adjacent non-cancerous tissues and benign fibroadenoma tissues (?2=43,P
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Objective To promote the migratory ability and immunological effect of bone marrow-derived dendritic cells ( BMDC) loaded with breast carcinoma antigen. Methods DCs were cultured by the medium containing rmGM-CSF and rmIL-4. After loaded with breast carcinoma antigen, DCs were stimulated with PGE2 for 1day. CD86, CD80, and CCR7 were measured by flow cytometry. The expression of CCR7 on surface of BMDC was also detected by RT-PCR and Western blotting. The chemotaxis assay was measured by migration through a polycarbonate filter in transwell chambers. The competence of inducing mixed lymphocyte response (MLR) and specific cytotoxic T lymphocyte ( CTL) were detected with MTT. The effect of DC blocking tumor growth in breast carcinoma model were also studied. Results Compared with control group, PGE2 upregulated surface markers of CD86, CD80, and CCR7 (P