PRELIMINARY STUDY ON THE MECHANISM OF CCL5/RANTES IN DIABETIC PATIENTS WITH DIFFUSE LARGE B CELL LYMPHOMA
(Abstract release date: 05/21/15)
EHA Library. zhang J. 06/12/15; 102672; PB1934
Disclosure(s): Jinhua Hospital of Zhejiang UniversityHematology
Jingcheng zhang
Contributions
Contributions
Abstract
Abstract: PB1934
Type: Publication Only
Background
The chemokine RANTES/CCL5 is an inducible, secreted inflammatory cytokine of small molecular weight, and has functions of chemotaxis and activation of T cells and mononuclear cell. The protein among them, containing 68 amino acid residues and its molecular weight being 8kd, is mainly formed by NK cells and T lymphocytes, and plays an important part in immunity to the tissue damage, infection and tumor.
Aims
The purpose of this experiment is to investigate the mechanism of chemokine CCL5 in diabetes mellitus with diffuse large B cell lymphoma (DLBCL), and to investigate the action of CCL5 gene from molecular level, cell level and animal level and partial molecular mechanism, in order to provide experimental data with reference value for the function mechanism of CCL5 in diabetes with DLBCL.
Methods
Normal human B cells and DLBCL cells were cultured in vitro, and RT-PCR was used to detect the expression of CCL5 mRNA; human diffuse large B lymphoma cell lines and rat diffuse large B cell lymphoma cell lines A20 were cultured in two kinds of sugar concentration of 5mmol/L and 30mmol/L, RT-PCR was applied for the detection of the expression of CCL5 mRNA respectively; BALB/ c mice were intraperitoneal injected streptozotocin (STZ) of small dose to construct diabetic rats model, and cell lines with a stable low expression of CCL5 and high expression of CCL5 were established via lentiviral transfection technology. The three kinds of cell lines of low expression of CCL5, high expression of CCL5 and un-transfection were injected subcutaneously in the diabetic BALB/c mice and normal blood glucose BALB/c mice, then were observed about the rate and the time of tumor formation and tumor size and texture; the expression of CCL5 in each group was detected by tumor tissue routine HE staining and immunohistochemistry; peripheral blood from mice in each group was extracted to detect the expression of CCL5 in peripheral blood by ELISA.
Results
1. DLBCL cell lines of Ly1, Ly8, Ly10 are cultured in vitro, the expression level of CCL mRNA in each group is higher than the normal B cell(P<0.05); 2. Glu30mmol/L cultivate human DLBCL cell strain Ly1, Ly8, Ly10 and mice DLBCL cell strain A20, after culturing for 2W, the CCL5mRNA are all higher than the one cultured by Glu5mmol/L?P<0.05?; 3. Through injecting the diabetic mice by lower expression CCL5, high expression CCL5, none transfection cell lines A20, the diabetic mice’s tumor formation rate is A1: 93.3%; A2: 60%; A3: 66.6%, the tumor forming time are respectively A1: 7.0±0.85d; A2: 9.5±2.8d; A3: 9.0±1.8d. Higher than the normal blood sugar mice’s tumor forming rate B1?20%?B2: 20%; B3: 46.6%, the tumor forming time are respectively B1: 12±1.3d; B2: 14±2.5d; B3: 12 ±4.2d; 4. Using the immuno-histochemistry to detect the diabetic mice tumor forming tissue CCL5’s expression, and the result is higher than the normal blood sugar mice’s tumor tissue;5. The result of ELISA detecting the diabetes mice’s CCL5 expression is higher than the mice with normal blood sugar in the peripheral blood.
Summary
1. Human DLBCL cell strain’s expression of CCL5mRNA are higher than the normal B cell; 2. High concentrated glucose cultivate human DLBCL cell strain and mice DLBCL cell strain’s CCL5mRNA is higher than the low sugar cultivate one, this shows in the high glucose environment, the DLBCL tumor cell can produce more CCL5; 3. The high expression CCL5’s cell strain is more easily to form tumor in the body than the low expression CCL5’s cell strain, the tumor grow faster; and at the same condition, the diabetes mice are more easily to form the tumor than the normal mice; 4. The expression of CCL5 in the tumor forming tissue of the diabetic mice is higher than the tumor forming tissue of the normal mice; 5. The expression of CCL5 in the serum of the diabetic mice is higher than the normal mice.
Type: Publication Only
Background
The chemokine RANTES/CCL5 is an inducible, secreted inflammatory cytokine of small molecular weight, and has functions of chemotaxis and activation of T cells and mononuclear cell. The protein among them, containing 68 amino acid residues and its molecular weight being 8kd, is mainly formed by NK cells and T lymphocytes, and plays an important part in immunity to the tissue damage, infection and tumor.
Aims
The purpose of this experiment is to investigate the mechanism of chemokine CCL5 in diabetes mellitus with diffuse large B cell lymphoma (DLBCL), and to investigate the action of CCL5 gene from molecular level, cell level and animal level and partial molecular mechanism, in order to provide experimental data with reference value for the function mechanism of CCL5 in diabetes with DLBCL.
Methods
Normal human B cells and DLBCL cells were cultured in vitro, and RT-PCR was used to detect the expression of CCL5 mRNA; human diffuse large B lymphoma cell lines and rat diffuse large B cell lymphoma cell lines A20 were cultured in two kinds of sugar concentration of 5mmol/L and 30mmol/L, RT-PCR was applied for the detection of the expression of CCL5 mRNA respectively; BALB/ c mice were intraperitoneal injected streptozotocin (STZ) of small dose to construct diabetic rats model, and cell lines with a stable low expression of CCL5 and high expression of CCL5 were established via lentiviral transfection technology. The three kinds of cell lines of low expression of CCL5, high expression of CCL5 and un-transfection were injected subcutaneously in the diabetic BALB/c mice and normal blood glucose BALB/c mice, then were observed about the rate and the time of tumor formation and tumor size and texture; the expression of CCL5 in each group was detected by tumor tissue routine HE staining and immunohistochemistry; peripheral blood from mice in each group was extracted to detect the expression of CCL5 in peripheral blood by ELISA.
Results
1. DLBCL cell lines of Ly1, Ly8, Ly10 are cultured in vitro, the expression level of CCL mRNA in each group is higher than the normal B cell(P<0.05); 2. Glu30mmol/L cultivate human DLBCL cell strain Ly1, Ly8, Ly10 and mice DLBCL cell strain A20, after culturing for 2W, the CCL5mRNA are all higher than the one cultured by Glu5mmol/L?P<0.05?; 3. Through injecting the diabetic mice by lower expression CCL5, high expression CCL5, none transfection cell lines A20, the diabetic mice’s tumor formation rate is A1: 93.3%; A2: 60%; A3: 66.6%, the tumor forming time are respectively A1: 7.0±0.85d; A2: 9.5±2.8d; A3: 9.0±1.8d. Higher than the normal blood sugar mice’s tumor forming rate B1?20%?B2: 20%; B3: 46.6%, the tumor forming time are respectively B1: 12±1.3d; B2: 14±2.5d; B3: 12 ±4.2d; 4. Using the immuno-histochemistry to detect the diabetic mice tumor forming tissue CCL5’s expression, and the result is higher than the normal blood sugar mice’s tumor tissue;5. The result of ELISA detecting the diabetes mice’s CCL5 expression is higher than the mice with normal blood sugar in the peripheral blood.
Summary
1. Human DLBCL cell strain’s expression of CCL5mRNA are higher than the normal B cell; 2. High concentrated glucose cultivate human DLBCL cell strain and mice DLBCL cell strain’s CCL5mRNA is higher than the low sugar cultivate one, this shows in the high glucose environment, the DLBCL tumor cell can produce more CCL5; 3. The high expression CCL5’s cell strain is more easily to form tumor in the body than the low expression CCL5’s cell strain, the tumor grow faster; and at the same condition, the diabetes mice are more easily to form the tumor than the normal mice; 4. The expression of CCL5 in the tumor forming tissue of the diabetic mice is higher than the tumor forming tissue of the normal mice; 5. The expression of CCL5 in the serum of the diabetic mice is higher than the normal mice.
Abstract: PB1934
Type: Publication Only
Background
The chemokine RANTES/CCL5 is an inducible, secreted inflammatory cytokine of small molecular weight, and has functions of chemotaxis and activation of T cells and mononuclear cell. The protein among them, containing 68 amino acid residues and its molecular weight being 8kd, is mainly formed by NK cells and T lymphocytes, and plays an important part in immunity to the tissue damage, infection and tumor.
Aims
The purpose of this experiment is to investigate the mechanism of chemokine CCL5 in diabetes mellitus with diffuse large B cell lymphoma (DLBCL), and to investigate the action of CCL5 gene from molecular level, cell level and animal level and partial molecular mechanism, in order to provide experimental data with reference value for the function mechanism of CCL5 in diabetes with DLBCL.
Methods
Normal human B cells and DLBCL cells were cultured in vitro, and RT-PCR was used to detect the expression of CCL5 mRNA; human diffuse large B lymphoma cell lines and rat diffuse large B cell lymphoma cell lines A20 were cultured in two kinds of sugar concentration of 5mmol/L and 30mmol/L, RT-PCR was applied for the detection of the expression of CCL5 mRNA respectively; BALB/ c mice were intraperitoneal injected streptozotocin (STZ) of small dose to construct diabetic rats model, and cell lines with a stable low expression of CCL5 and high expression of CCL5 were established via lentiviral transfection technology. The three kinds of cell lines of low expression of CCL5, high expression of CCL5 and un-transfection were injected subcutaneously in the diabetic BALB/c mice and normal blood glucose BALB/c mice, then were observed about the rate and the time of tumor formation and tumor size and texture; the expression of CCL5 in each group was detected by tumor tissue routine HE staining and immunohistochemistry; peripheral blood from mice in each group was extracted to detect the expression of CCL5 in peripheral blood by ELISA.
Results
1. DLBCL cell lines of Ly1, Ly8, Ly10 are cultured in vitro, the expression level of CCL mRNA in each group is higher than the normal B cell(P<0.05); 2. Glu30mmol/L cultivate human DLBCL cell strain Ly1, Ly8, Ly10 and mice DLBCL cell strain A20, after culturing for 2W, the CCL5mRNA are all higher than the one cultured by Glu5mmol/L?P<0.05?; 3. Through injecting the diabetic mice by lower expression CCL5, high expression CCL5, none transfection cell lines A20, the diabetic mice’s tumor formation rate is A1: 93.3%; A2: 60%; A3: 66.6%, the tumor forming time are respectively A1: 7.0±0.85d; A2: 9.5±2.8d; A3: 9.0±1.8d. Higher than the normal blood sugar mice’s tumor forming rate B1?20%?B2: 20%; B3: 46.6%, the tumor forming time are respectively B1: 12±1.3d; B2: 14±2.5d; B3: 12 ±4.2d; 4. Using the immuno-histochemistry to detect the diabetic mice tumor forming tissue CCL5’s expression, and the result is higher than the normal blood sugar mice’s tumor tissue;5. The result of ELISA detecting the diabetes mice’s CCL5 expression is higher than the mice with normal blood sugar in the peripheral blood.
Summary
1. Human DLBCL cell strain’s expression of CCL5mRNA are higher than the normal B cell; 2. High concentrated glucose cultivate human DLBCL cell strain and mice DLBCL cell strain’s CCL5mRNA is higher than the low sugar cultivate one, this shows in the high glucose environment, the DLBCL tumor cell can produce more CCL5; 3. The high expression CCL5’s cell strain is more easily to form tumor in the body than the low expression CCL5’s cell strain, the tumor grow faster; and at the same condition, the diabetes mice are more easily to form the tumor than the normal mice; 4. The expression of CCL5 in the tumor forming tissue of the diabetic mice is higher than the tumor forming tissue of the normal mice; 5. The expression of CCL5 in the serum of the diabetic mice is higher than the normal mice.
Type: Publication Only
Background
The chemokine RANTES/CCL5 is an inducible, secreted inflammatory cytokine of small molecular weight, and has functions of chemotaxis and activation of T cells and mononuclear cell. The protein among them, containing 68 amino acid residues and its molecular weight being 8kd, is mainly formed by NK cells and T lymphocytes, and plays an important part in immunity to the tissue damage, infection and tumor.
Aims
The purpose of this experiment is to investigate the mechanism of chemokine CCL5 in diabetes mellitus with diffuse large B cell lymphoma (DLBCL), and to investigate the action of CCL5 gene from molecular level, cell level and animal level and partial molecular mechanism, in order to provide experimental data with reference value for the function mechanism of CCL5 in diabetes with DLBCL.
Methods
Normal human B cells and DLBCL cells were cultured in vitro, and RT-PCR was used to detect the expression of CCL5 mRNA; human diffuse large B lymphoma cell lines and rat diffuse large B cell lymphoma cell lines A20 were cultured in two kinds of sugar concentration of 5mmol/L and 30mmol/L, RT-PCR was applied for the detection of the expression of CCL5 mRNA respectively; BALB/ c mice were intraperitoneal injected streptozotocin (STZ) of small dose to construct diabetic rats model, and cell lines with a stable low expression of CCL5 and high expression of CCL5 were established via lentiviral transfection technology. The three kinds of cell lines of low expression of CCL5, high expression of CCL5 and un-transfection were injected subcutaneously in the diabetic BALB/c mice and normal blood glucose BALB/c mice, then were observed about the rate and the time of tumor formation and tumor size and texture; the expression of CCL5 in each group was detected by tumor tissue routine HE staining and immunohistochemistry; peripheral blood from mice in each group was extracted to detect the expression of CCL5 in peripheral blood by ELISA.
Results
1. DLBCL cell lines of Ly1, Ly8, Ly10 are cultured in vitro, the expression level of CCL mRNA in each group is higher than the normal B cell(P<0.05); 2. Glu30mmol/L cultivate human DLBCL cell strain Ly1, Ly8, Ly10 and mice DLBCL cell strain A20, after culturing for 2W, the CCL5mRNA are all higher than the one cultured by Glu5mmol/L?P<0.05?; 3. Through injecting the diabetic mice by lower expression CCL5, high expression CCL5, none transfection cell lines A20, the diabetic mice’s tumor formation rate is A1: 93.3%; A2: 60%; A3: 66.6%, the tumor forming time are respectively A1: 7.0±0.85d; A2: 9.5±2.8d; A3: 9.0±1.8d. Higher than the normal blood sugar mice’s tumor forming rate B1?20%?B2: 20%; B3: 46.6%, the tumor forming time are respectively B1: 12±1.3d; B2: 14±2.5d; B3: 12 ±4.2d; 4. Using the immuno-histochemistry to detect the diabetic mice tumor forming tissue CCL5’s expression, and the result is higher than the normal blood sugar mice’s tumor tissue;5. The result of ELISA detecting the diabetes mice’s CCL5 expression is higher than the mice with normal blood sugar in the peripheral blood.
Summary
1. Human DLBCL cell strain’s expression of CCL5mRNA are higher than the normal B cell; 2. High concentrated glucose cultivate human DLBCL cell strain and mice DLBCL cell strain’s CCL5mRNA is higher than the low sugar cultivate one, this shows in the high glucose environment, the DLBCL tumor cell can produce more CCL5; 3. The high expression CCL5’s cell strain is more easily to form tumor in the body than the low expression CCL5’s cell strain, the tumor grow faster; and at the same condition, the diabetes mice are more easily to form the tumor than the normal mice; 4. The expression of CCL5 in the tumor forming tissue of the diabetic mice is higher than the tumor forming tissue of the normal mice; 5. The expression of CCL5 in the serum of the diabetic mice is higher than the normal mice.
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