BACKGROUND Leukemia stem cells(LSCs)are found to be one of the main factors contributing to poor therapeutic effects in acute myeloid leukemia(AML),as they are protected by the bone marrow microenvironment(BMM)against...BACKGROUND Leukemia stem cells(LSCs)are found to be one of the main factors contributing to poor therapeutic effects in acute myeloid leukemia(AML),as they are protected by the bone marrow microenvironment(BMM)against conventional therapies.Gossypol acetic acid(GAA),which is extracted from the seeds of cotton plants,exerts anti-tumor roles in several types of cancer and has been reported to induce apoptosis of LSCs by inhibiting Bcl2.AIM To investigate the exact roles of GAA in regulating LSCs under different microenvironments and the exact mechanism.METHODS In this study,LSCs were magnetically sorted from AML cell lines and the CD34+CD38-population was obtained.The expression of leucine-rich pentatricopeptide repeat-containing protein(LRPPRC)and forkhead box M1(FOXM1)was evaluated in LSCs,and the effects of GAA on malignancies and mitochondrial RESULTS LRPPRC was found to be upregulated,and GAA inhibited cell proliferation by degrading LRPPRC.GAA induced LRPPRC degradation and inhibited the activation of interleukin 6(IL-6)/janus kinase(JAK)1/signal transducer and activator of transcription(STAT)3 signaling,enhancing chemosensitivity in LSCs against conventional chemotherapies,including L-Asparaginase,Dexamethasone,and cytarabine.GAA was also found to downregulate FOXM1 indirectly by regulating LRPPRC.Furthermore,GAA induced reactive oxygen species accumulation,disturbed mitochondrial homeostasis,and caused mitochondrial dysfunction.By inhibiting IL-6/JAK1/STAT3 signaling via degrading LRPPRC,GAA resulted in the elimination of LSCs.Meanwhile,GAA induced oxidative stress and subsequent cell damage by causing mitochondrial damage.CONCLUSION Taken together,the results indicate that GAA might overcome the BMM protective effect and be considered as a novel and effective combination therapy for AML.展开更多
目的:研究联合中性粒细胞百分比(Percentage of neutrophils,NEU%)、C-反应蛋白(C-reactive protein,CRP)及白介素-6(Interleukin-6,IL-6)在白血病合并肺部感染中的诊断价值。方法:选取2019年1月到2023年1月在我院收治的81例白血病患者...目的:研究联合中性粒细胞百分比(Percentage of neutrophils,NEU%)、C-反应蛋白(C-reactive protein,CRP)及白介素-6(Interleukin-6,IL-6)在白血病合并肺部感染中的诊断价值。方法:选取2019年1月到2023年1月在我院收治的81例白血病患者作为研究对象。根据感染标准分为未感染组(36例)和感染组(45例)。感染组患者进一步根据感染程度分级,分为轻度感染组(32例)与重度感染组(13例)。对比未感染组和感染组外周血中NEU%、CRP、IL-6水平;对比轻度感染组与重度感染组外周血中NEU%、CRP、IL-6水平;探讨NEU%、CRP、IL-6水平单独及联合检测对白血病合并感染的诊断效能。结果:对比两组外周血中NEU%、CRP、IL-6水平,感染组3个指标均高于未感染组(P<0.05)。轻度感染组中NEU%、CPR、IL-6指标水平均低于重度感染组(P<0.05)。NEU%特异度为50.31%,灵敏度为80.20%,准确度为69.83%,CRP特异度为50.00%,灵敏度为88.37%,准确度为75.69%,IL-6特异度为40.36%,灵敏度为92.32%,准确度为72.32%,三项联合特异度为67.32%,灵敏度为96.51%,准确度为86.62%(P<0.05)。结论:白血病合并肺部感染中联合NEU%、CRP及IL-6水平明显升高,且三者联合对白血病合并肺部感染诊断价值高。展开更多
There is currently no effective targeted therapeutic strategy for the treatment of central nervous system acute lymphoblastic leukemia(CNS-ALL).Integrinα6 is considered a potential target for CNS-ALL diagnosis and th...There is currently no effective targeted therapeutic strategy for the treatment of central nervous system acute lymphoblastic leukemia(CNS-ALL).Integrinα6 is considered a potential target for CNS-ALL diagnosis and therapy because of its role in promoting CNS-ALL disease progression.The targeted peptide D(RWYD)(abbreviated RD),with nanomolar affinity to integrinα6 was identified by peptide scanning techniques such as alanine scanning,truncation,and D-substitution.Herein,we developed a therapeutic nanoparticle based on the integrinα6-targeted peptide for treating CNS-ALL.The self-assembled proapoptotic nanopeptide_(D)(RWYD)-_(D)(KLAKLAK)_(2)-G_(D)(FFY)(abbreviated RD-KLA-Gffy)contains the integrinα6-targeted peptide RD,the well-known proapoptotic peptide_(D)(KLAKLAK)_(2)(abbreviated KLA),and the self-assembling tetrapeptide GD(FFY)(abbreviated Gffy).The functional mechanism of RD-KLA-Gffy is clarified using different experiments.Our results demonstrate that RD-KLA-Gffy is highly enriched in CNS-ALL lesions and induces tumor cell apoptosis,thus reducing CNS-ALL disease burden and prolonging the survival of CNS-ALL mice without obvious toxicity.Moreover,the combined use of RD-KLA-Gffy and methotrexate(MTX)shows a potent antitumor effect in treating CNS-ALL,indicating that RD-KLA-Gffy plays an important role in suppressing CNS-ALL progression either as a single agent or in combination with MTX,which shows promise for application in CNS-ALL therapy.展开更多
Acute myeloid leukemia(AML) is a heterogeneous disease characterized by the accu.mulation of immature myeloid progenitor cells in the bone marrow,compromising of normal hematopoi.esis and ultimately resulting in bone ...Acute myeloid leukemia(AML) is a heterogeneous disease characterized by the accu.mulation of immature myeloid progenitor cells in the bone marrow,compromising of normal hematopoi.esis and ultimately resulting in bone marrow failure.Chemotherapy is the mainstay treatment for all AML patients,however,drug resistance and clinical relapse limits its efficacy.The 5-year survival rate of AML patients is only 26.6%.Survival rates are even lower among patients ages 65 to 74 years(5.3%) and 75 years or older(1.6%).Therefore,exploring novel therapeutic agents is urgent for improving the outcome of patients with AML.Saponins are amphipathic glycosides found in traditional Chinese medicines.In the present study,we isolated a panel of saponins from Paris forrestii(Takht.) H.Li,a unique plant found in Tibet and Yunnan provinces,China.By examining their activities in suppressing acute myeloid leukemia cell proliferation,total saponins from Paris forrestii(TSPf) displayed more potent activity than individual ones.TSPf induced more than 40% AML cell apoptosis within 24 h and decreased the viability of all leukemia cell lines.TSPf-induced apoptosis was confirmed by both Annexin V staining and caspase-3 activation.TSPf downregulated pro-survival proteins Mcl-1,Bcl-xL and Bcl-2,but upreg.ulated the expression of tumor suppressor proteins p53,p27,Bax and Beclin 1.The AKT/mTOR signaling pathway is frequently over activated in various AML cells,and TSPf was found to suppress the activa.tion of both AKT and mTOR,but had no effects on their total protein expression.This was further con.firmed by the inactivation of 4 EBP-1 and p70 S6 K,two typical downstream signal molecules in the AKT/mTOR pathway.More specifically,TSPf-inactivated AKT/mTOR signaling was found to be associated with downregulated RNF6,a recently identified oncogene in AML.RNF6 activated AKT/mTOR,and consistently,knockdown of RNF6 led to inactivation of the AKT/mTOR pathway.Furthermore,TSPf suppressed the growth of AML xenografts in nude mice models.Oral administration of 100 mg · kg^(-1) body weight almost fully suppressed tumor growth within 14 d,without gross toxicity.This study thus demonstrated that TSPf displays potent anti-AML activity by suppressing the RNF6/AKT/mTOR pathway.Given its low toxicity,TSPf could be developed for the treatment of AML.展开更多
This work was supported by a grant from the National Natural Science Foundation of China (No.39730420). This is one of papers of the special issue on gene therapy research (Chin J Cancer Res Vol. 9 No. 4 December, 19...This work was supported by a grant from the National Natural Science Foundation of China (No.39730420). This is one of papers of the special issue on gene therapy research (Chin J Cancer Res Vol. 9 No. 4 December, 1997). Human IL 6 gene was transduced into FBL 3 murine erythroleukemia cells in vitro by calcium phos phate co participation. After selection in the presence of G418, limiting dilution and biological activity assay, G418 resistant clone that secreted the highest level of IL 6 (225.6 U/ml) was selected out of 24 IL 6 secreting clones. The FBL 3 cells secreting the highest level of IL 6 (FBL 3 IL 6) showed decreased growth potential and clono genicity in vitro. Inhibition of cell growth and clone formation was found to be closely related to the level of IL 6 secretion. FBL 3 IL 6 cells grew more slowly than wild type FBL 3 leukemia cells and FBL 3 cells secreting lower level of IL 6 (21.3 U/ml) when inoculated s.c. into C57BL/6 mice. The mice inoculated with FBL 3 IL 6 cells showed prolonged survival period than those inoculated with control leukemia cells. Increased cytotoxic activities of splenic NK and CTL were found in mice inoculated with FBL 3 IL 6 cells. The secretions of IL 2, TNF and GM CSF from murine splenocytes were also found to be greatly elevated after the inoculation of FBL 3 IL 6 leukemia cells. These data suggested that transduction of IL 6 gene into FBL 3 cells magnificently decreased the tumorigenicity and increased the immunogenicity of the leukemia cells, could induce specific and nonspecific antitumor immune responses. IL 6 gene modified leukemia cells might be of great interests to be used as vaccine for the treatment of leukemia.展开更多
Objective: To investigate the antitumor effect of the IL 6 gene modified erythroleukemia cells combined with low dose cyclophosphamide (Cy) and low dose IL 2 Methods: Mice inoculated with FBL 3 IL 6 in combin...Objective: To investigate the antitumor effect of the IL 6 gene modified erythroleukemia cells combined with low dose cyclophosphamide (Cy) and low dose IL 2 Methods: Mice inoculated with FBL 3 IL 6 in combination with low dose IL 2 and low dose cyclophosphamide (Cy) Results: Mice received combined therapy of FBL 3 IL 6, IL 2 and Cy developed tumors most slowly and survived much longer when compared with mice in control groups, with 5 out of 8 leukemia bearing mice being tumor free 100 days after the combined treatment To further explain the mechanism of the antitumor effects by the combined therapy It was found that combined therapy with low dose Cy, low dose IL 2 and FBL 3 IL 6 achieved maximal cytotoxic effects of nature killer cells and specific cytotoxic T lymphocytes, increased production IL 2, TNF and GM CSF from spleen lymphocytes in tumor bearing mice Vaccination with the FBL3 IL 6 also enhanced the cytotoxic activity of the peritoneal macrophages The results demonstrated that administration of low dose Cy and low dose IL 2 in combination with IL 6 gene modified leukemia vaccine could elicit potent anti leukemia effects, and the mechanisms involved in the antitumor process may include the induction of specific and nonspecific antitumor immunity, reversal of T suppressor cells that mediated local immuno suppression in tumor bearing mice Conclusion: The combined therapy with cytokine gene modified tumor vaccine, low dose of Cy and IL 2 might be a promising approach for the treatment of leukemia展开更多
Objective: To evaluate the expression and its clinical significance of interleukin 6 (IL-6), soluble glycoprotein 130 (sgp130), interleukin 8 (IL-8) and type A interleukin 8 receptor (IL-8RA) in acute promyelocytic le...Objective: To evaluate the expression and its clinical significance of interleukin 6 (IL-6), soluble glycoprotein 130 (sgp130), interleukin 8 (IL-8) and type A interleukin 8 receptor (IL-8RA) in acute promyelocytic leukemia (APL) patients during all-trans retinoic acid (ATRA) induction treatment. Methods: Plasma and bone marrow mononuclear cell (MNC) culture supernatant IL-6, sgp130, IL-8 concentration of 18 cases with APL were kinetically measured in vivo and in vitro (ELISA). Bone marrow MNC IL-8RA was measured by flow cytometry after being cultured with ATRA (10?6mmol/L). Results: Plasma IL-6, sgp130, IL-8 levels were higher than normal (P<0.05), IL-6, spg130 levels correlated with white blood cell (WBC) counts (P<0.05) while IL-8 levels correlated with body temperature (P<0.05) at initial diagnosis. After 72-hour incubation with ATRA, concentration of IL-6 of bone marrow MNC culture supernatant did not change, that of sgp130 mildly decreased, and IL-8 significantly decreased while the positive rate of IL-8RA on bone marrow MNC increased. During ATRA treatment, plasma IL-6 changes were correlated with WBC counts. Peak levels of IL-6 and WBC were lower in patients who received intermittent therapy than those who received continuous therapy. Plasma IL-6 and IL-8 were increased when complicated with infection and IL-8 seemed more sensitive. Conclusion: Plasma IL-6, sgp130, IL-8 levels may reflect patients' responsiveness to ATRA treatment, and could be used to predict hyperleukocytosis and intercurrent infection. ATRA induces APL cell differentiation possibly via sgp130 signal transducer chain. Measurement of sgp130 had certain meaning to prognosis.展开更多
Mouse FBL-3 erythroleukemia cells were transfected with recombinant human interleukin-6 (rhIL-6) gene and a clone secreting IL-6 was selected (i.e., FBL-3-IL-6+). The cell clone secreting IL-6 was inoculated into C57B...Mouse FBL-3 erythroleukemia cells were transfected with recombinant human interleukin-6 (rhIL-6) gene and a clone secreting IL-6 was selected (i.e., FBL-3-IL-6+). The cell clone secreting IL-6 was inoculated into C57BL/6 mouse. The growth of tumors was observed and histologic analyses of the tumors in situ, liver, spleen and bone marrow were performed after inoculation. The mice inoculated with wild-type FBL-3 erythroleukemia cells were used as the control. The results showed that the later the tumor occurrence, the slower the tumor development,the lower the pathological changes degree and the longer the survival time in experimental group compared to that of the control. The results demonstrated that the inoculation of the FBL-3 cell clone secreting IL-6 can inhibit the invasion of leukemia cells, suggesting that the FBL-3-IL-6+ cells can he used as a vaccine to treat leukemia.展开更多
Objective We aimed to evaluate the expression pattern of the genes BIM, BCL-6, and c-MYC in adult patients at initial diagnosis of B-cell acute lymphoblastic leukemia(B-ALL).Methods Relative m RNA levels of BIM, BCL-6...Objective We aimed to evaluate the expression pattern of the genes BIM, BCL-6, and c-MYC in adult patients at initial diagnosis of B-cell acute lymphoblastic leukemia(B-ALL).Methods Relative m RNA levels of BIM, BCL-6, and c-MYC in peripheral blood mononuclear cells(PBMCs) from B-ALL patients were determined by quantitative reverse-transcription polymerase chain reaction(q RT-PCR) using SYBR Green dye. PBMCs from healthy volunteers served as a control. GAPDH was used as a reference gene.Results Relative expression of BIM, BCL-6, and c-MYC m RNA in B-ALL patients was significantly lower than in healthy controls(P < 0.05). Furthermore, this result was observed for both newly diagnosed B-ALL patients and those incomplete remission(CR)(P < 0.05). There were no statistically significant differences in the expression levels of BIM, BCL-6, and c-MYC between these B-ALL patient groups(P > 0.05). Spearman's rank correlation analyses revealed the expression level of BIM to be positively correlated with that of BCL-6 in B-ALL patients.Conclusion Expression of the genes BIM, BCL-6, and c-MYC is decreased in adult B-ALL patients. Moreover, the expression pattern of these genes may be similar in such patients at initial diagnosis and following CR. The expression characteristics of BIM, BCL-6, and c-MYC may constitute useful markers for the diagnosis of adult B-ALL.展开更多
Objective：To investigate the association between the X-ray repair cross complementing（XRCC） group 5, XRCC6 and XRCC7 polymorphisms and risk of acute myeloid leukemia（AML）. Methods：This hospital-based case-contro...Objective：To investigate the association between the X-ray repair cross complementing（XRCC） group 5, XRCC6 and XRCC7 polymorphisms and risk of acute myeloid leukemia（AML）. Methods：This hospital-based case-control study included 120 AML patients and 210 cancer-free controls in a Chinese population. Three polymorphisms of XRCC5, XRCC6 and XRCC7 were genotyped using the polymerase chain reaction（PCR） or polymerase chain reaction-restriction fragment length polymorphism（PCR-RFLP） method. Results： We found that there was a significant decrease in risk of AML associated with the XRCC6 -61 CG/GG genotype（adjusted odd ratio （OR） = 0.55; 95% confident interval（CI） = 0.34-0.89） compared with the -61CC genotype. For the novel tandem repeat polymorphism （VNTR） in the XRCC5 promoter, we found when the XRCC5 six genotypes were dichotomized（i.e., 2R/2R, 2R/1R versus 2R/0R, 1R/1R, 1R/0R and 0R/0R）, the latter group was associated with increased risk of AML（adjusted OR = 1.67; 95% CI = 1.00-2.79） compared to 2R/ 2R＋2R/1R genotype. However, the XRCC7 6721G〉T polymorphism had no effect on risk of AML. Conclusion：The XRCC6 -61C 〉 G and XRCC5 2R/1R/0R polymorphisms, but not XRCC7 6721G 〉 T polymorphism, could play an important role in the development of AML. Larger scale studies with more detailed data on environment exposure are needed to verify these findings.展开更多
文摘BACKGROUND Leukemia stem cells(LSCs)are found to be one of the main factors contributing to poor therapeutic effects in acute myeloid leukemia(AML),as they are protected by the bone marrow microenvironment(BMM)against conventional therapies.Gossypol acetic acid(GAA),which is extracted from the seeds of cotton plants,exerts anti-tumor roles in several types of cancer and has been reported to induce apoptosis of LSCs by inhibiting Bcl2.AIM To investigate the exact roles of GAA in regulating LSCs under different microenvironments and the exact mechanism.METHODS In this study,LSCs were magnetically sorted from AML cell lines and the CD34+CD38-population was obtained.The expression of leucine-rich pentatricopeptide repeat-containing protein(LRPPRC)and forkhead box M1(FOXM1)was evaluated in LSCs,and the effects of GAA on malignancies and mitochondrial RESULTS LRPPRC was found to be upregulated,and GAA inhibited cell proliferation by degrading LRPPRC.GAA induced LRPPRC degradation and inhibited the activation of interleukin 6(IL-6)/janus kinase(JAK)1/signal transducer and activator of transcription(STAT)3 signaling,enhancing chemosensitivity in LSCs against conventional chemotherapies,including L-Asparaginase,Dexamethasone,and cytarabine.GAA was also found to downregulate FOXM1 indirectly by regulating LRPPRC.Furthermore,GAA induced reactive oxygen species accumulation,disturbed mitochondrial homeostasis,and caused mitochondrial dysfunction.By inhibiting IL-6/JAK1/STAT3 signaling via degrading LRPPRC,GAA resulted in the elimination of LSCs.Meanwhile,GAA induced oxidative stress and subsequent cell damage by causing mitochondrial damage.CONCLUSION Taken together,the results indicate that GAA might overcome the BMM protective effect and be considered as a novel and effective combination therapy for AML.
文摘目的:研究联合中性粒细胞百分比(Percentage of neutrophils,NEU%)、C-反应蛋白(C-reactive protein,CRP)及白介素-6(Interleukin-6,IL-6)在白血病合并肺部感染中的诊断价值。方法:选取2019年1月到2023年1月在我院收治的81例白血病患者作为研究对象。根据感染标准分为未感染组(36例)和感染组(45例)。感染组患者进一步根据感染程度分级,分为轻度感染组(32例)与重度感染组(13例)。对比未感染组和感染组外周血中NEU%、CRP、IL-6水平;对比轻度感染组与重度感染组外周血中NEU%、CRP、IL-6水平;探讨NEU%、CRP、IL-6水平单独及联合检测对白血病合并感染的诊断效能。结果:对比两组外周血中NEU%、CRP、IL-6水平,感染组3个指标均高于未感染组(P<0.05)。轻度感染组中NEU%、CPR、IL-6指标水平均低于重度感染组(P<0.05)。NEU%特异度为50.31%,灵敏度为80.20%,准确度为69.83%,CRP特异度为50.00%,灵敏度为88.37%,准确度为75.69%,IL-6特异度为40.36%,灵敏度为92.32%,准确度为72.32%,三项联合特异度为67.32%,灵敏度为96.51%,准确度为86.62%(P<0.05)。结论:白血病合并肺部感染中联合NEU%、CRP及IL-6水平明显升高,且三者联合对白血病合并肺部感染诊断价值高。
基金supported by grants from the National Natural Science Foundation of China (81972531, 82373175, 82102775, and 82002466)the Major Scientific and Technological Projects of Guangdong Province (2019B020202002)the Young Talents Program of Sun Yat-sen University Cancer Center (YTP-SYSUCC-0067)
文摘There is currently no effective targeted therapeutic strategy for the treatment of central nervous system acute lymphoblastic leukemia(CNS-ALL).Integrinα6 is considered a potential target for CNS-ALL diagnosis and therapy because of its role in promoting CNS-ALL disease progression.The targeted peptide D(RWYD)(abbreviated RD),with nanomolar affinity to integrinα6 was identified by peptide scanning techniques such as alanine scanning,truncation,and D-substitution.Herein,we developed a therapeutic nanoparticle based on the integrinα6-targeted peptide for treating CNS-ALL.The self-assembled proapoptotic nanopeptide_(D)(RWYD)-_(D)(KLAKLAK)_(2)-G_(D)(FFY)(abbreviated RD-KLA-Gffy)contains the integrinα6-targeted peptide RD,the well-known proapoptotic peptide_(D)(KLAKLAK)_(2)(abbreviated KLA),and the self-assembling tetrapeptide GD(FFY)(abbreviated Gffy).The functional mechanism of RD-KLA-Gffy is clarified using different experiments.Our results demonstrate that RD-KLA-Gffy is highly enriched in CNS-ALL lesions and induces tumor cell apoptosis,thus reducing CNS-ALL disease burden and prolonging the survival of CNS-ALL mice without obvious toxicity.Moreover,the combined use of RD-KLA-Gffy and methotrexate(MTX)shows a potent antitumor effect in treating CNS-ALL,indicating that RD-KLA-Gffy plays an important role in suppressing CNS-ALL progression either as a single agent or in combination with MTX,which shows promise for application in CNS-ALL therapy.
基金supported by Natural Science Foundation of China(81770154)
文摘Acute myeloid leukemia(AML) is a heterogeneous disease characterized by the accu.mulation of immature myeloid progenitor cells in the bone marrow,compromising of normal hematopoi.esis and ultimately resulting in bone marrow failure.Chemotherapy is the mainstay treatment for all AML patients,however,drug resistance and clinical relapse limits its efficacy.The 5-year survival rate of AML patients is only 26.6%.Survival rates are even lower among patients ages 65 to 74 years(5.3%) and 75 years or older(1.6%).Therefore,exploring novel therapeutic agents is urgent for improving the outcome of patients with AML.Saponins are amphipathic glycosides found in traditional Chinese medicines.In the present study,we isolated a panel of saponins from Paris forrestii(Takht.) H.Li,a unique plant found in Tibet and Yunnan provinces,China.By examining their activities in suppressing acute myeloid leukemia cell proliferation,total saponins from Paris forrestii(TSPf) displayed more potent activity than individual ones.TSPf induced more than 40% AML cell apoptosis within 24 h and decreased the viability of all leukemia cell lines.TSPf-induced apoptosis was confirmed by both Annexin V staining and caspase-3 activation.TSPf downregulated pro-survival proteins Mcl-1,Bcl-xL and Bcl-2,but upreg.ulated the expression of tumor suppressor proteins p53,p27,Bax and Beclin 1.The AKT/mTOR signaling pathway is frequently over activated in various AML cells,and TSPf was found to suppress the activa.tion of both AKT and mTOR,but had no effects on their total protein expression.This was further con.firmed by the inactivation of 4 EBP-1 and p70 S6 K,two typical downstream signal molecules in the AKT/mTOR pathway.More specifically,TSPf-inactivated AKT/mTOR signaling was found to be associated with downregulated RNF6,a recently identified oncogene in AML.RNF6 activated AKT/mTOR,and consistently,knockdown of RNF6 led to inactivation of the AKT/mTOR pathway.Furthermore,TSPf suppressed the growth of AML xenografts in nude mice models.Oral administration of 100 mg · kg^(-1) body weight almost fully suppressed tumor growth within 14 d,without gross toxicity.This study thus demonstrated that TSPf displays potent anti-AML activity by suppressing the RNF6/AKT/mTOR pathway.Given its low toxicity,TSPf could be developed for the treatment of AML.
文摘This work was supported by a grant from the National Natural Science Foundation of China (No.39730420). This is one of papers of the special issue on gene therapy research (Chin J Cancer Res Vol. 9 No. 4 December, 1997). Human IL 6 gene was transduced into FBL 3 murine erythroleukemia cells in vitro by calcium phos phate co participation. After selection in the presence of G418, limiting dilution and biological activity assay, G418 resistant clone that secreted the highest level of IL 6 (225.6 U/ml) was selected out of 24 IL 6 secreting clones. The FBL 3 cells secreting the highest level of IL 6 (FBL 3 IL 6) showed decreased growth potential and clono genicity in vitro. Inhibition of cell growth and clone formation was found to be closely related to the level of IL 6 secretion. FBL 3 IL 6 cells grew more slowly than wild type FBL 3 leukemia cells and FBL 3 cells secreting lower level of IL 6 (21.3 U/ml) when inoculated s.c. into C57BL/6 mice. The mice inoculated with FBL 3 IL 6 cells showed prolonged survival period than those inoculated with control leukemia cells. Increased cytotoxic activities of splenic NK and CTL were found in mice inoculated with FBL 3 IL 6 cells. The secretions of IL 2, TNF and GM CSF from murine splenocytes were also found to be greatly elevated after the inoculation of FBL 3 IL 6 leukemia cells. These data suggested that transduction of IL 6 gene into FBL 3 cells magnificently decreased the tumorigenicity and increased the immunogenicity of the leukemia cells, could induce specific and nonspecific antitumor immune responses. IL 6 gene modified leukemia cells might be of great interests to be used as vaccine for the treatment of leukemia.
文摘Objective: To investigate the antitumor effect of the IL 6 gene modified erythroleukemia cells combined with low dose cyclophosphamide (Cy) and low dose IL 2 Methods: Mice inoculated with FBL 3 IL 6 in combination with low dose IL 2 and low dose cyclophosphamide (Cy) Results: Mice received combined therapy of FBL 3 IL 6, IL 2 and Cy developed tumors most slowly and survived much longer when compared with mice in control groups, with 5 out of 8 leukemia bearing mice being tumor free 100 days after the combined treatment To further explain the mechanism of the antitumor effects by the combined therapy It was found that combined therapy with low dose Cy, low dose IL 2 and FBL 3 IL 6 achieved maximal cytotoxic effects of nature killer cells and specific cytotoxic T lymphocytes, increased production IL 2, TNF and GM CSF from spleen lymphocytes in tumor bearing mice Vaccination with the FBL3 IL 6 also enhanced the cytotoxic activity of the peritoneal macrophages The results demonstrated that administration of low dose Cy and low dose IL 2 in combination with IL 6 gene modified leukemia vaccine could elicit potent anti leukemia effects, and the mechanisms involved in the antitumor process may include the induction of specific and nonspecific antitumor immunity, reversal of T suppressor cells that mediated local immuno suppression in tumor bearing mice Conclusion: The combined therapy with cytokine gene modified tumor vaccine, low dose of Cy and IL 2 might be a promising approach for the treatment of leukemia
文摘Objective: To evaluate the expression and its clinical significance of interleukin 6 (IL-6), soluble glycoprotein 130 (sgp130), interleukin 8 (IL-8) and type A interleukin 8 receptor (IL-8RA) in acute promyelocytic leukemia (APL) patients during all-trans retinoic acid (ATRA) induction treatment. Methods: Plasma and bone marrow mononuclear cell (MNC) culture supernatant IL-6, sgp130, IL-8 concentration of 18 cases with APL were kinetically measured in vivo and in vitro (ELISA). Bone marrow MNC IL-8RA was measured by flow cytometry after being cultured with ATRA (10?6mmol/L). Results: Plasma IL-6, sgp130, IL-8 levels were higher than normal (P<0.05), IL-6, spg130 levels correlated with white blood cell (WBC) counts (P<0.05) while IL-8 levels correlated with body temperature (P<0.05) at initial diagnosis. After 72-hour incubation with ATRA, concentration of IL-6 of bone marrow MNC culture supernatant did not change, that of sgp130 mildly decreased, and IL-8 significantly decreased while the positive rate of IL-8RA on bone marrow MNC increased. During ATRA treatment, plasma IL-6 changes were correlated with WBC counts. Peak levels of IL-6 and WBC were lower in patients who received intermittent therapy than those who received continuous therapy. Plasma IL-6 and IL-8 were increased when complicated with infection and IL-8 seemed more sensitive. Conclusion: Plasma IL-6, sgp130, IL-8 levels may reflect patients' responsiveness to ATRA treatment, and could be used to predict hyperleukocytosis and intercurrent infection. ATRA induces APL cell differentiation possibly via sgp130 signal transducer chain. Measurement of sgp130 had certain meaning to prognosis.
文摘Mouse FBL-3 erythroleukemia cells were transfected with recombinant human interleukin-6 (rhIL-6) gene and a clone secreting IL-6 was selected (i.e., FBL-3-IL-6+). The cell clone secreting IL-6 was inoculated into C57BL/6 mouse. The growth of tumors was observed and histologic analyses of the tumors in situ, liver, spleen and bone marrow were performed after inoculation. The mice inoculated with wild-type FBL-3 erythroleukemia cells were used as the control. The results showed that the later the tumor occurrence, the slower the tumor development,the lower the pathological changes degree and the longer the survival time in experimental group compared to that of the control. The results demonstrated that the inoculation of the FBL-3 cell clone secreting IL-6 can inhibit the invasion of leukemia cells, suggesting that the FBL-3-IL-6+ cells can he used as a vaccine to treat leukemia.
基金Supported by the gants of the Guangdong Province Key Foundation of Science and Technology Program(No.2009B0507000029)the Major Research Plan of the National Natural Science Foundation of China(cultivating project,No.91129720)+1 种基金the Guangdong Science&Technology Project(Nos.2012B050600023 and 2015A050502029)a Grant from the Overseas Chinese Affairs Office of the State Council Key Discipline Construction Fund(No.51205002)
文摘Objective We aimed to evaluate the expression pattern of the genes BIM, BCL-6, and c-MYC in adult patients at initial diagnosis of B-cell acute lymphoblastic leukemia(B-ALL).Methods Relative m RNA levels of BIM, BCL-6, and c-MYC in peripheral blood mononuclear cells(PBMCs) from B-ALL patients were determined by quantitative reverse-transcription polymerase chain reaction(q RT-PCR) using SYBR Green dye. PBMCs from healthy volunteers served as a control. GAPDH was used as a reference gene.Results Relative expression of BIM, BCL-6, and c-MYC m RNA in B-ALL patients was significantly lower than in healthy controls(P < 0.05). Furthermore, this result was observed for both newly diagnosed B-ALL patients and those incomplete remission(CR)(P < 0.05). There were no statistically significant differences in the expression levels of BIM, BCL-6, and c-MYC between these B-ALL patient groups(P > 0.05). Spearman's rank correlation analyses revealed the expression level of BIM to be positively correlated with that of BCL-6 in B-ALL patients.Conclusion Expression of the genes BIM, BCL-6, and c-MYC is decreased in adult B-ALL patients. Moreover, the expression pattern of these genes may be similar in such patients at initial diagnosis and following CR. The expression characteristics of BIM, BCL-6, and c-MYC may constitute useful markers for the diagnosis of adult B-ALL.
基金supported in part by National Natural Science Foundation of China(30571541)Natural Science Foundation of Jiangsu Province(BK2006233,BK2005161)+1 种基金Medicine Foundation of Jiangsu Province(H200506)Creative Science Foundation of Nanjing Medical University(CX2004002)
文摘Objective：To investigate the association between the X-ray repair cross complementing（XRCC） group 5, XRCC6 and XRCC7 polymorphisms and risk of acute myeloid leukemia（AML）. Methods：This hospital-based case-control study included 120 AML patients and 210 cancer-free controls in a Chinese population. Three polymorphisms of XRCC5, XRCC6 and XRCC7 were genotyped using the polymerase chain reaction（PCR） or polymerase chain reaction-restriction fragment length polymorphism（PCR-RFLP） method. Results： We found that there was a significant decrease in risk of AML associated with the XRCC6 -61 CG/GG genotype（adjusted odd ratio （OR） = 0.55; 95% confident interval（CI） = 0.34-0.89） compared with the -61CC genotype. For the novel tandem repeat polymorphism （VNTR） in the XRCC5 promoter, we found when the XRCC5 six genotypes were dichotomized（i.e., 2R/2R, 2R/1R versus 2R/0R, 1R/1R, 1R/0R and 0R/0R）, the latter group was associated with increased risk of AML（adjusted OR = 1.67; 95% CI = 1.00-2.79） compared to 2R/ 2R＋2R/1R genotype. However, the XRCC7 6721G〉T polymorphism had no effect on risk of AML. Conclusion：The XRCC6 -61C 〉 G and XRCC5 2R/1R/0R polymorphisms, but not XRCC7 6721G 〉 T polymorphism, could play an important role in the development of AML. Larger scale studies with more detailed data on environment exposure are needed to verify these findings.
