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Objective:To investigate the risk factors for overall survival in operable hepatocellular carcinoma with portal vein tumor thrombus (PVTT-HCC) patients and establish a scoring system.Methods:Survival data in 253 PVTT-HCC patients were retrospectively analyzed in Guangxi Medical University Affiliated Tumor Hospital. Survival curves were analyzed using the Kaplan-Meier method and log-rank test. Cox stepwise regression analysis was used to identify independent preoperative risk factors affecting overall survival. A prognostic scoring system based on independent risk factors and their relative coefficients was established to screen patients with greater hepatic resection benefits, and the identification ability of the model was based on ROC.Results:A total of 253 patients with PVTT-HCC were enrolled in this study, there were 222 males and 31 females, with a median age 44 years. The median survival time in all patients was (13.00±2.15) months. Rate of overall survival was 51.8% at 1 year, 25.0% at 3 years and 17.7% at 5 years. Multivariable Cox regression analyses showed four risk factors including: AST≥40 U/L, ALP (≥80 U/L), tumor number (>1), and incomplete tumor capsule. A prognostic scoring system was established based on these variables. The area under curve of the scoring system was 0.780 (95% CI: 0.715-0.845). Patients were classified as low- or high-risk group for hepatic resection depending on whether their score was <3 ( n=77) or ≥3 ( n=176), respectively. High-risk patients had a median survival of 10 months, compared to 29 months in low-risk patients. Low-risk patients also had better survival rates at 1 year (75.3% vs 41.5%), 3 years (47.6% vs 15.2%), and 5 years (34.7% vs 10.5%), P<0.05. Conclusion:A prognostic scoring system for hepatic resection in PVTT-HCC patients has been developed based entirely on preoperative variables. Using this system, patients belong to the low risk group have better prognosis after surgery, which can provide a basis for surgical treatment of PVTT-HCC patients.
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Objective@#To compare the prognosis of radiofrequency ablation (RFA) for postoperative recurrent hepatocellular carcinoma and primary hepatocellular carcinoma(HCC).@*Methods@#The clinical data of 179 patients with recurrent HCC (recurrent group) and primary HCC (primary group) treated by RFA from 2009 to 2015 were retrospectively analyzed. Overall survival rate (OS) and disease-free survival rate (DFS) were analyzed by Kaplan-meier log-rank test. The prognostic factors of RFA for recurrent HCC were analyzed by COX proportional hazard regression.@*Results@#The 1, 3 and 5year′s OS of the recurrent group were 93%, 73%, 61%, respectively and 85%, 75%, 61% for the primary group(χ2=0.017, P=0.896). The corresponding 1, 3 and 5year′s DFS were 61%, 39%, 21% and 79%, 64%, 46% respectively (χ2=3.899, P=0.048). The independent risk factors affecting the OS of the recurrent group were the interval between hepatectomy to recurrence≤12 months (HR=0.264, 95% CI=0.077-0.901, P=0.033) and the Child-Pugh grading B before RFA (HR=4.501, 95% CI=1.426-14.208, P=0.01).@*Conclusions@#The DFS of patients with recurrent HCC were shorter than that with primary HCC treated by RFA. The interval between hepatectomy to recurrence and the Child-Pugh grading before RFA were independent risk factors for OS of the recurrent group.
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Tet2 (member 2 of the Tet family) plays an important role in DNA demethylation modification, epigenetic regulation, and hematopoiesis. In our previous study, we found that Tet2 knockout mice progressively developed lymphocytic leukemia and myeloid leukemia with aging. However，the role of Tet2 in bone marrow microenvironment is unclear. Here in this study, we found that more Tet2-/- mesenchymal stem cells (MSCs) from bone marrow were in the G2/M cell cycle stages. The division time of Tet2-/- MSCs was shorter than that of the control cells. The growth rate of Tet2-/- MSCs was accelerated. The cobblestone area-forming cells assay (CAFC) showed that Tet2 knockout MSCs supported the expansion of hematopoietic stem cells (HSCs) and the differentiation of HSCs was skewed towards myeloid cells. Through the dot blotting experiment, we found that the total methylation level was increased in Tet2-/- bone marrow cells (BM). We used the methylation-chip to analyze the methylation level of Tet2-/- bone marrow cells and found that the level of methylation was increased in the transcriptional starting area (TSS), exons (EXONS) and 3' untranslated region (3' UTR). Moreover, we found that the cytokines secreted by Tet2-/- MSCs, such as IL-8 and IL-18, were decreased. While the expressions of GM-CSF and CCL-3, which supported hematopoietic stem cells to differentiate to myeloid cells, were increased in Tet2-/- MSCs. Our results demonstrated that Tet2 regulates MSCs to support hematopoiesis.
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Animals , Mice , Bone Marrow Cells , Cell Differentiation , DNA-Binding Proteins , Epigenesis, Genetic , Hematopoiesis , Hematopoietic Stem Cells , Mesenchymal Stem Cells , Proto-Oncogene ProteinsABSTRACT
Objective To compare the prognosis of radiofrequency ablation (RFA) for postoperative recurrent hepatocellular carcinoma and primary hepatocellular carcinoma(HCC).Methods The clinical data of 179 patients with recurrent HCC (recurrent group) and primary HCC (primary group) treated by RFA from 2009 to 2015 were retrospectively analyzed.Overall survival rate (OS) and disease-free survival rate (DFS) were analyzed by Kaplan-meier log-rank test.The prognostic factors of RFA for recurrent HCC were analyzed by COX proportional hazard regression.Results The 1,3 and 5year's OS of the recurrent group were 93％,73％,61％,respectively and 85％,75％,61％ for the primary group(x2 =0.017,P =0.896).The corresponding 1,3 and 5year's DFS were 61％,39％,21％ and 79％,64％,46％ respectively (x2 =3.899,P =0.048).The independent risk factors affecting the OS of the recurrent group were the interval between hepatectomy to recurrence ≤ 12 months (HR =0.264,95％ CI =0.077-0.901,P =0.033) and the Child-Pugh grading B before RFA (HR =4.501,95％ CI =1.426-14.208,P =0.01).Conclusions The DFS of patients with recurrent HCC were shorter than that with primary HCC treated by RFA.The interval between hepatectomy to recurrence and the Child-Pugh grading before RFA were independent risk factors for OS of the recurrent group.
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Objective To establish a preoperative nomogram model in predicting microvascular invasion (MVI) and to test its predictive effectiveness in hepatocellular carcinoma (HCC).Methods This retrospective study was conducted on 798 patients with HCC,including 690 males and 108 females,aged (49.8± 10.9) years old who underwent curative hepatectomy in the Guangxi Medical University Affiliated Tumor Hospital between January 2014 and December 2017 were retrospectively analyzed.The patients were divided into the model group (n=579) and the validation group (n=219) according to the periods of the operation time.Independent risk factors of MVI were identified by univariate and multivariate logistic regression analysis in the model group,and a nomogram model was established according to the independent risk factors.The accuracy of the nomogram model in predicting MVI was detected in the two groups by the computer consistency coefficient (C-index) and calibration graph method.The predictive value was evaluated by receiver operating characteristic curve.Results Histopathological diagnosis revealed 278 patients with MVI and no MVI in the 301 patients of HCC out of the 579 patients in the model group.In the validation group,there were 119 patients with MVI and 100 patients with no MVI out of the 219 patients.Total bilirubin ＞15 μmol/L(OR=1.519,95％ CI:1.041 ～ 2.217),alkaline phosphatase ＞60 U/L(OR =1.681,95％CI:1.059～2.670),alpha-fetoprotein ＞200 ng/L (OR=2.192,95％CI:1.531 ～3.134) and tumor maximum diameter (OR =1.120,95％CI:1.057 ～ 1.187) were the independent risk factors of MVI on multivariate analysis.After establishment of the nomogram model using the independent risk factors,the C-indexes were 0.680 and 0.773 respectively in the model group and the validation group.In the calibration graph,the standard curve properly fitted with the predicting calibration curve.The predicted value of MVI obtained was in good agreement with the observed value.The ROC curve analysis nomogram model predicted the low performance of MVI.Conclusion The nomogram model in predicting MVI in patients with HCC was successfully established.The model offered certain guiding significance in the clinical treatment of HCC.
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Objective To retrospectively study the prognostic impact of paraneoplastic erythrocytosis (PE) on patients with hepatocellular carcinoma (HCC) after liver resection.Methods 713 patients with HCC who underwent partial hepatic resection in The Affiliated Cancer Hospital of Guangxi Medical University were divided into two groups:the PE group (n =81) and the non-PE group (n =632).The overall survival between the two groups were compared after reducing confounding bias by using propensity score matching (PSM).Independent prognostic predictors were determined by the Cox proportional hazards model.Results 80 pairs of patients were matched using PSM.In the matched cohort,the PE group exhibited significantly longer overall survival (OS) compared to the NPE group of patients without erythrocytosis.The 1-,3-,and 5-year overall survival rates were 88.6％,74.2％,69.0％ in the PE group,and 91.0％,60.1％,41.6％ in the non-PE group,respectively (P ＜ 0.05).Using the log-rank test,tumor size ≥ 10cm,macrovascular invasion,Barcelona Clinic Liver Cancer (BCLC) stage C,PE and complete tumor encapsulation were significantly associated with OS in patients with HCC after liver resection.The Cox regression analysis indicated that tumor size ≥ 10 cm,and Barcelona Clinic Liver Cancer (BCLC) stage C were independent prognostic factors of poor prognosis,while complete tumor encapsulation and paraneoplastic erythrocytosis were independent predictors of good prognosis.Conclusions For patients with HCC who underwent surgical resection,patients with PE had better prognosis than those without PE under the condition of similar tumor burden.PE was an independent predictor of good prognosis.
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Objective@#To explore the role of PDK1 in the transition of endothelial to hematopoietic cells and its effect on the generation and normal function of HSC.@*Methods@#PDK1 was deleted specifically in endothelial cells expressing VEC (Vascular Endothelial Cadherin). CFU-C was performed to detect the effect of PDK1 on the function of hematopoietic progenitor cells using the cells from PDK1fl/fl, PDK1fl/+ and Vec-Cre; PDK1fl/fl AGM region. Hematopoietic stem cell transplantation assay was conducted to determine the effect of PDK1 on hematopoietic stem cells. Flow cytometry was performed to analyze the influence of PDK1 on percentage, cell cycle and apoptosis of CD31+c-Kithigh cell population. Real-time PCR was conducted to measure the expression of transcription factors involved in process of transition from endothelial to hematopoietic cells.@*Results@#In contrast to the wild type group, the CFU from PDK1-deficient hematopoietic progenitor cells showed smaller in morphology and fewer in quantity. CFU-GM was (24±5)/ee in knockout group, and the control group was (62±1)/ee (P=0.001). PDK1 deletion severely impaired the ability to repopulate hematopoietic cells and differentiate into committed cells. hematopoietic progenitor cells from knockout group was transplanted into 5 recipients without any recipients reconstructed. However, 5 of 7 recipients were reconstructed in control group (P=0.001). The proportion of intra-vascular clusters in the AGM was decreased (the frequency of CD31+c-Kithigh in the knockout group was (0.145±0.017)%, and the control group ratio was (0.385±0.040)% (P=0.001), but not due to the inhibition of cell proliferation and/or increase of apoptosis. Further study found that the absence of endothelial PDK1 causes a decreased expression of RUNX1, P2-RUNX1, GATA2 and other important hematopoietic-related transcription factors in hemogenic cluster.@*Conclusion@#PDK1 deletion impairs the transition of endothelial cells to hematopoietic cells as well as the generation and function of HSC.
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Objective To compare the clinical features and surgical outcomes of hepatocellular carcinoma (HCC) combined with portal venous tumor thrombus (PVTT) and hepatic venous tumor thrombus (HVTT) or bile duct tumor thrombi (BDTT),and analyze the effects of different tumor thrombus (TT) types and different surgical methods for TT on prognosis.Methods The retrospective cross-sectional study was conducted.The clinical data of 220 HCC patients with lymphovascular invasion (LVI) who were admitted to the Affiliated Cancer Hospital of Guangxi Medical University between January 2004 and December 2014 were collected.Of 220 patients,140 were combined with PVTT,36 with HVTT and 44 with BDTT.According to patients' conditions,they underwent tumor and TT resection,and tumor resection + TT removal or single TT removal.Observation indicators:(1) comparisons of clinical features of HCC patients with PVTT or HVTT or BDTT;(2) surgical and postoperative situations;(3) follow-up and survival.Follow-up using outpatient examination and telephone interview was performed to detect postoperative survival up to December 2015.Measurement data with normal distribution were represented as (x)±s.Comparisons among 3 indicators were analyzed using the one-way ANOVA,and comparisons between 2 indicators were analyzed using the t test.Comparisons of count data were analyzed using the chi-square test.The survival curve and rate were respectively drawn and calculated by the Kaplan-Meier method,and the Log-rank test was used for survival analysis.Results (1) Comparisons of clinical features of HCC patients with PVTT or HVTT or BDTT:number of patients with Child-pugh A,Child-pugh B and peritoneal effusion,tumor diameter and cases with tumor capsule were respectively detected in 133,7,23,(10±4)cm,91 in HCC patients with PVTT and 35,1,4,(9±4)cm,27 in HCC patients with HVTT and 35,9,16,(6±4)cm,15 in HCC patients with BDTT,with statistically significant differences (x2 =12.693,10.408,F=11.300,x2 =17.188,P＜ 0.05).(2) Surgical and postoperative situations:of 140 HCC patients with PVTT,51 underwent tumor and PVTT resection,89 underwent tumor resection + PVTT removal through incising portal vein;68 received postoperative transcatheter arterial chemoembolization (TACE).Thirty-six HCC patients with HVTT underwent tumor and HVTT resection;24 received postoperative TACE.Of 44 HCC patients with BDTT,23 underwent tumor and BDTT resection,21 underwent tumor resection + BDTT removal through incising common bile duct;29 received postoperative TACE.(3) Follow-up and survival:① 220 patients were followed up for 1-73 months,with a median time of 12 months.The median survival time,1-,3-and 5-year survival rates were respectively 12 months,48.2％,25.0％,15.4％ in 140 HCC patients with PVTT and 28 months,77.1％,45.6％,24.5％ in 36 HCC patients with HVTT and 36 months,88.6％,48.3％,24.6％ in 44 HCC patients with BDTT,with a statistically significant difference in survival (x2 =13.316,P＜0.05).② Of 140 HCC patients with PVTT,49 were in type Ⅰ PVTT,and median survival time,1-,3-and 5-year survival rates were respectively 20 months,60.3％,32.6％ and 17.1％;70 were in type Ⅱ PVTT,and median survival time,1-,3-and 5-year survival rates were respectively 13 months,51.4％,26.0％ and 17.3％;21 were in type Ⅲ PVTT,and median survival time,1-,3-and 5-year survival rates were respectively 7 months,9.5％,4.8％ and 0,showing a statistically significant difference in survival (x2=18.102,P＜0.05).The median survival time,1-,3-and 5-year survival rates were respectively 21 months,72.5％,42.5％,26.2％ in 51 patients undergoing tumor and TT resection and 9 months,40％,14.4％,0 in 89 patients undergoing tumor resection + PVTT removal through incising portal vein,showing a statistically significant difference in survival (x2=24.098,P＜0.05).③ Of 36 HCC patients with HVTT,17 were detected in right HVTT,and median survival time,1-,3-and 5-year survival rates were respectively 14 months,64.7％,20.2％ and 0;10 were detected in left HVTT,and median survival time,1-,3-and 5-year survival rates were respectively 53 months,80.0％,70.0％ and 38.9％;9 were detected in middle HVTT,and median survival time,1-,3-and 5-year survival rates were respectively 40 months,88.9％,61.0％ and 30.5％;showing no statistically significant difference in survival (x2 =5.951,P＞0.05).④ Of 44 HCC patients with BDTT,24,6 and 14 were respectively detected in type Ⅰ,Ⅱ and Ⅲ BDTTs,and median survival time,1-,3-and 5-year survival rates were respectively 38 months,87.5％,60.4％,34.9％ in type Ⅰ BDTT patients and 26 months,83.3％,16.7％,0 in type Ⅱ BDTT patients and 35 months,78.6％,50.0％,21.4％ in type Ⅲ BDTT patients,showing no statistically significant difference in survival (x2 =5.312,P＞0.05).Of 44 patients,median survival time,1-,3-and 5-year survival rates were respectively 38 months,91.3％,59.5％,34.3％ in 23 patients undergoing tumor and TT resection and 26 months,85.7％,35.7％,15.3％ in 21 patients undergoing tumor resection + TT removal through incising common bile duct,showing no statistically significant difference in survival (x2 =2.071,P＞0.05).Conclusions HCC patients with PVTT have larger tumor diameter and worse liver dysfunction,and are prone to peritoneal effusion.HCC patients with different LVI undergo surgery.There is better prognosis in HCC patients with BDTT,and good prognosis in patients with HVTT,while poorer prognosis in patients with PVTT.The postoperative survival of HCC patients with PVTT is associated with TT type,and patients will have better prognosis after tumor resection + TT removal if TT type is confirmed earlier.The postoperative survival of HCC patients with BDTT is not associated with TT type,tumor resection + TT removal maybe prolong postoperative survival time.
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As a dioxygenase, Ten-Eleven Translocation 2 (TET2) catalyzes subsequent steps of 5-methylcytosine (5mC) oxidation. TET2 plays a critical role in the self-renewal, proliferation, and differentiation of hematopoietic stem cells, but its impact on mature hematopoietic cells is not well-characterized. Here we show that Tet2 plays an essential role in osteoclastogenesis. Deletion of Tet2 impairs the differentiation of osteoclast precursor cells (macrophages) and their maturation into bone-resorbing osteoclasts in vitro. Furthermore, Tet2 mice exhibit mild osteopetrosis, accompanied by decreased number of osteoclasts in vivo. Tet2 loss in macrophages results in the altered expression of a set of genes implicated in osteoclast differentiation, such as Cebpa, Mafb, and Nfkbiz. Tet2 deletion also leads to a genome-wide alteration in the level of 5-hydroxymethylcytosine (5hmC) and altered expression of a specific subset of macrophage genes associated with osteoclast differentiation. Furthermore, Tet2 interacts with Runx1 and negatively modulates its transcriptional activity. Our studies demonstrate a novel molecular mechanism controlling osteoclast differentiation and function by Tet2, that is, through interactions with Runx1 and the maintenance of genomic 5hmC. Targeting Tet2 and its pathway could be a potential therapeutic strategy for the prevention and treatment of abnormal bone mass caused by the deregulation of osteoclast activities.
Subject(s)
Animals , Mice , 5-Methylcytosine , Chemistry , Metabolism , Cell Differentiation , Cells, Cultured , Core Binding Factor Alpha 2 Subunit , Genetics , Metabolism , DNA-Binding Proteins , Physiology , Genome , Genomics , Mice, Knockout , Osteoclasts , Cell Biology , Metabolism , Proto-Oncogene Proteins , PhysiologyABSTRACT
Objective To investigate the role of perioperative antiviral therapy with Entecavir for patients with hepatitis B virus related hepatocellular carcinoma (HBV-HCC) with low serum HBV DNA levels.Methods The HVB-HCC patients were randomly divided into 2 groups.Patients in the antiviral group received Entecavir 4 days before hepatic resection while patients in the control group received no antiviral treatment.The serum HBV DNA,liver function,morbidity and length of hospital stay were compared between the two groups.Results Sixteen patients in the control group (n =44) developed HBV reactivation.On the other hand,only 1 patient in the antiviral group (n =44) developed HBV reactivation.Recovery in liver function in the antiviral group was much faster than the control group,especially for glutamic-pyruvic transaminase level.The antiviral group had significantly lower morbidity and shorter total or postoperative hospitalization (all P ＜ 0.05).Conclusions Patients with HBV-HCC with low levels of HBV DNA have a high risk of developing HBV reactivation in the perioperative period.Perioperative antiviral therapy was safe and efficacious in preventing HBV reactivation,improved liver function,reduced postoperative complications and shortened hospitalization.
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To study the biological function of DNAH2 (Homo sapiens dynein, axonemal, heavy chain 2) gene, we constructed human stable U2OS cell line of DNAH2 gene knockout through CRISPR/Cas9n double nick system. The A, B sgRNAs (Single guide RNA) and complementary strands were designed and synthesized. The double-stranded structures were formed during annealing, and connected with BbsⅠ cohesive ends-containing pX462 linear vector to construct the recombinant eukaryotic expression plasmids, including pX462-DNAH2-A and pX462-DNAH2-B. After the co-transfection of the two plasmids into U2OS cells, the addition of puromycin and limiting dilution method were used to obtain positive monoclonal cell line. Western blotting assay was then performed to detect the expression of DNAH2 protein, and PCR-sequencing technology was finally utilized to analyze the mutation feature. The results showed that A, B sgRNAs duplex was successfully inserted into pX462 vector, and DNAH2 protein was not expressed and DNAH2 gene suffered from the frame-shift mutation in U2OS-DNAH2-KO monoclonal cell line. These demonstrated that DNAH2 knockout U2OS stable cell line was successfully constructed through CRISPR/Cas9n double nick system, which providing a useful tool for the study of DNAH2 gene.
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Objective To prepare specific mouse monoclonal antibodies against Homo sapiens dynein,axonemal, heavy chain 2 (DNAH2). Methods Firstly, recombinant plasmid encoding His tagged immunogen, targeting N-terminal sequence of DNAH2 protein (1-300 aa), in E. coli was constructed. IPTG was used to induce the expression of His-immunogen, which was then purified and immunized in BALB/c mice. Hybridoma cells were obtained through the fusion between myeloma cells and splenocytes isolated from BALB/c mice. Finally, ELISA and Western blot assays were performed to screen the positive hybridoma. Results IPTG was used efficiently to induce the expression of DNAH2 immunogen in E. coli. DNAH2 protein bands were detected in screened positive hybridoma. Conclusion Mouse monoclonal anti-DNAH2 antibody is prepared successfully.
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<p><b>OBJECTIVE</b>To establish the ADAR1 (adenosine deaminase that act on RNA 1) knockout MLL-AF9 acute myeloid leukemia (AML) mouse model, and to preliminarily investigate the effects of ADAR1 deletion on the development of AML.</p><p><b>METHODS</b>The lineage⁻ (Lin⁻) cells of ER-CreADAR1(lox/lox) mice and their ADAR1(lox/lox) counterparts were enriched by magnetic activated cell sorting (MACS) and then transduced with retrovirus carrying MSCV- MLL/AF9-IRES-GFP fusion gene. The efficiency of transduction was detected by flow cytometry, and equal number of GFP⁺ cells were transplanted into lethally irradiated recipient mice. The recipient mice were treated with tamoxifen at 48 hours after transplantation to induce ADAR1 knockout and divided into following groups: experimental group (ER-Cre;ADAR1(lox/lox)+tamoxifen), control groups ((1)ER-Cre;ADAR1(lox/lox)+vechile, (2)ADAR1(lox/lox)+tamoxifen, (3)ADAR1(lox/lox)+vechile). The percentage of GFP⁺ cells in peripheral blood was examined at 10, 15 and 20 days respectively after transplantation and the survival of the recipient mice was observed. In vitro study, ER-Cre;ADAR1(lox/lox) and ADAR1(lox/lox) AML cells were cultured and the apoptosis rates of these cells 48 hours after 4-hydroxytamoxifen treatment were examined.</p><p><b>RESULTS</b>The ADAR1 deletion MLL-AF9 AML mouse model was successfully established. Deletion of ADAR1 could decrease the percentage of GFP⁺ cells in the peripheral blood and significantly prolong the survival rate of recipient mice（P<0.05）. In vitro study showed that the cultured total cell number, percentage of GFP⁺ cells decreased and the apoptosis rate of AML cells increased.</p><p><b>CONCLUSION</b>Ablation of ADAR1 could delay the progression of AML in recipient mice. ADAR1 plays a critical role in the development and maintenance of murine MLL-AF9 AML.</p>
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Animals , Mice , Adenosine Deaminase , Apoptosis , Disease Models, Animal , Leukemia, Myeloid, Acute , Myeloid-Lymphoid Leukemia Protein , TamoxifenABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of PD0332991 (C24H29N7O2) on cell cycle, apoptosis, differentiation and self-renewal of hematopoietic stem cells (HSC) in mice.</p><p><b>METHODS</b>The self renewal ability of HSCs was measured by cobblestone forming cell assay (CAFC). The colony-forming cell (CFC) assay was used to quantify the changes of numbers and functions of HPC after the treatment of the compound. The expressions of self-renewal regulation genes, cell cycle-related genes, apoptosis-related genes were measured by real-time PCR. The cell cycle status and apoptosis of HSC and HPC were analyzed by flow cytometry.</p><p><b>RESULTS</b>There were obvious changes in cell cycle regulation between control and PD0332991 groups. HSCs in G1 phase increased significantly from 76.3% to 89.5% after treatment of PD0332991 (P<0.05) while cells in S, G2 and M phase reduced from 20.5% to 7.3% (P<0.05). HPCs in G1 phase also increased from 74% to 87.4% after treatment of PD0332991 (P<0.05) while cells in S, G2 and M phase reduced from 25.54% to 11.6% (P<0.05). The apoptotic fractions between control and PD0332991 groups had no statistical difference (P>0.05). After cultured with PD0332991, the expression levels of cell cycle genes CDK1, CyclinA2, CyclinF, p18, p19 and p27 decreased by 58.77%, 66.35%, 56.33%, 62.18%, 32.28% and 36.53% respectively, while expression of CDK7 increased by 27.27% (P<0.05). No visible expression difference was observed in apoptosis and self-renew related genes. After treatment of PD0332991, the self-renewal ability of HSC decreased significantly. There were almost no CFCs in PD0332991 group in CAFC assay. Similarly, the frequency of CFCs was dramatically lower in PD0332991 group.</p><p><b>CONCLUSION</b>These results suggested that PD0332991 affected HSC/HPC from mice mainly through inhibiting the cell cycle rather than apoptosis. It also suggested that CDK4/6 might play a key role in the regulation of HSC/HPC.</p>
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Animals , Mice , Apoptosis , Cell Cycle , Cell Differentiation , Cell Proliferation , Cells, Cultured , Hematopoietic Stem Cells , Cell Biology , Mice, Inbred C57BL , Piperazines , Pharmacology , Pyridines , PharmacologyABSTRACT
The protein,the fats,the carbohydrate and the nucleic acid are the main constituent material of normal cell and organizations. In the early process of becoming malignant,these chemical substance's structure,the conformation and quantity has had the obvious change,but this time clinical symptoms and medicine phantom study still has not obvious change.But the Raman spectrum can actually reflect these changes.Therefore the Raman spectrum technology becomes important means of the malignant tumor early diagnosis and the cancer mechanism research hopefully.This article makes a summary on the present as well as the progress reviewed of Raman spectrum technology in the tumor research application.