CN102580059A - Application of thymosin extrasin alpha in preparing medicine for preventing and curing cancer of liver - Google Patents
Application of thymosin extrasin alpha in preparing medicine for preventing and curing cancer of liver Download PDFInfo
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- CN102580059A CN102580059A CN2012100861660A CN201210086166A CN102580059A CN 102580059 A CN102580059 A CN 102580059A CN 2012100861660 A CN2012100861660 A CN 2012100861660A CN 201210086166 A CN201210086166 A CN 201210086166A CN 102580059 A CN102580059 A CN 102580059A
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Abstract
Application of thymosin extrasin alpha in preparing medicines for preventing and curing cancer of the liver relates to thymosin extrasin alpha, and effect of the thymosin extrasin alpha in restaining regulatory T-lymphocyte cells is further provided. The effect of recombinant proteins in preventing and curing the cancers of the liver is observed by injecting the thymosin extrasin alpha in intraperitoneal mode, and size of the tumor of the liver and ascites forming prevention are included. Through injection treatment and prevention of the thymosin extrasin alpha, growth of the tumor of the liver can be obviously restrained and simultaneously formation of the ascites can be reduced. By observing level of the regulatory T-lymphocyte cells of mouse splenic organs, the level of the regulatory T-lymphocyte cells of splenic organs in a contrast group is proved to be obviously higher than that of regulatory T-lymphocyte cells of normal mouse splenic organs, and difference is remarkable (P<0.01); and the level of the regulatory T-lymphocyte cells of splenic organs in a medicine-used group is obviously lower than that of the normal contrast group and the experiment contrast group, and the difference is obvious (P<0.01).
Description
Technical field
The present invention relates to prophymosin-alpha, especially relate to the purposes of a kind of prophymosin-alpha in preparation prevention and treatment liver-cancer medicine.
Background technology
1984, Haritos etc. isolated a kind of polypeptide that contains 111 amino acid residues from rat chest gland, because its N end contains all prophymosin-alpha family members' that found sequence, so the called after prophymosin-alpha.People ProT α is made up of 109 amino acid residues, compares with the sequence of P of Rats roT α, and the difference in 6 sites is arranged; Comprise 4 sites substitute with the disappearance in 2 sites (1, Haritos AA; Goodall GJ, Horecker BL, Prothymosin alpha:isolation and properties of the major immunoreactive form of thymosin alphal in rat thymus.Proc Natl Acad Sci U S A; 1984,81:1008; 2, Haritos AA.Alpha-thymosins:relationships in structure, distribution, and function.Isozymes Curr Top Biol Med Res 1987,14:123-152; 3, Goodall GJ, Dominguez F, Horecker BL.Molecular cloning of cDNA for human prothymosin alpha.Proc Natl Acad Sci U S A 1986; 83 (23): 8926-8928; 4, Pan LX; Haritos AA, Wideman J, Komiyama T; Chang M, Stein S et al.Human prothymosin alpha:amino acid sequence and immunologic properties.Arch Biochem Biophys 1986; 250 (1): 197-201).
1985, Haritos etc. (5, Komiyama T, Pan LX, Haritos AA, et al.The primary structure of rat parathymosin.Proc Natl Acad Sci U S A 1986; 83 (5): 1242-1245) find that P of Rats roTA can strengthen the ability that mouse anti Candida albicans (Candida albicans) infects; Pan in 1986 finds that again ProT α can stimulate the release of tumor metastasis suppressor gene (MIF); Its effect than 10～20 times of T α the last 1 (6, Haritos AA; R.Blacher; S.Stein, J..Horecker.Parathymosin alpha:a peptide from rat tissues with structural homology to prothymosin alpha.Proc Natl Acad Sci U S A, 82:1050-1053; 7, Pan LX, Haritos AA, Wideman J, et al.Human prothymosin alpha:amino acid sequence and immunologic properties.Arch Biochem Biophys 1986; 250 (1): 197-201); 1987; Discovery lumbar injection ProT α such as S α lvin can strengthen the ability that the RF/J mice produces antibody; Zinc can be strengthened this effect, can make the cellular immunization of young rat and senile rat and humoral immunization all strengthen (8, Salvin SB, Horecker BL; Pan LX, Rabin BS.The effect of dietary zinc and prothymosin alpha on cellular immune responses of RF/J mice.Clin Immunol Immunopathol 1987; 43 (3): 281-288.)。Papanastasiou(9、Papanastasiou?M，Baxevanis?CN，Papamichail?M.Promotion?of?murine?antitumor?activity?by?prothymosin?alpha?treatment：I.Induction?of?tumoricidal?peritoneal?cells?producing?high?levels?of?tumour?necrosis?factor?alpha.Cancer?Immunol?Immunother?1992；35(2)：145-150。) find; If it is CBA/2 mice abdominal cavity inoculated tumour cell produces ascites within general 8-12 days, dead after 10-14 days; 20% the mice of handling through ProT α does not produce ascites, and can make the mouse life that does not produce the 40%-60% in the ascites can extend to 70 days.When lumbar injection ProT α; Can strengthen macrophage, improve N K; LA K cell activity (10, Lopez-Rodriguez JL; Cordero OJ, Sarandeses C .Interleukin-2 killer cells:in vitro evaluation of combination with prothymosin alpha.Lymphokine Cytokine Res 1994; 13 (3): 175-182; 11, Baxevanis CN, Gritzapis AD, Dedoussis GV .Induction of lymphokine-activated killer activity in mice by prothymosin alpha.Cancer Immunol Immunother 1994; 38 (4): 281-286); Strengthen the MHC restriction cellular immunization (12, Baxevanis CN; Thanos D; Reclos GJ, J et al.Prothymosin alpha enhances human and murine MHC class II surface antigen expression and messenger RNA accumulation.J Immunol 1992; 148 (7): 1979-1984; 13, Baxevanis CN; Thanos D; Reclos GJ, et al.Prothymosin alpha enhances human and murine MHC class II surface antigen expression and messenger RNA accumulation.J Immunol 1992; 148 (7): 1979-1984); Suppress systemic lupus erythematosus (sle) (14, Baxevanis CN; Reclos CJ; Papamichail M et al.Prothymosin alpha restores the depressed autologous and allogenic mixed lymphocyte responses in patient with systemic lupus erythematosus.Immuopharmaco immunotoxico, 1987,9:429.), promote IL-2, the secretion of MIF and TN F-α, IFN-γ's is synthetic; Promote the IL-2 receptor expression (15, Baxevanis CN; Gritzapis AD; Spanakos G .Induction of tumor-specific T lymphocyte responses in vivo by prothymosin alpha.Cancer Immunol Immunother 1995; 40 (6): 410-418; 16, Lopez-Rodriguez JL; Cordero OJ; Sarandeses C .Interleukin-2 killer cells:in vitro evaluation of combination with prothymosin alpha.Lymphokine Cytokine Res 1994; 13 (3): 175-182; 17, Baxevanis CN, Gritzapis AD, Dedoussis GV .Induction of lymphokine-activated killer activity in mice by prothymosin alpha.Cancer Immunol Immunother 1994; 38 (4): 281-286; 18, Voutsas IF; Baxevanis CN; Gritzapis AD, et al.Synergy between interleukin-2 and prothymosin alpha for the increased generation of cytotoxic T lymphocytes against autologous human carcinomas.Cancer Immunol Immunother 2000; 49 (8): 449-458; 19, Cordero OJ; Sarandeses CS, Lopez JL, et al.Prothymosin alpha enhances IL-2receptor expression in norma human T-lymphoctyes.Int J Immunpharmacol; 1991,13:1059.), thereby produce non-specific tumor-killing effect, simultaneously, the specificity antineoplastic effect of inducing tumor-specific cytotoxic T cell (CD8+) and helper T lymphocyte (CD4+).ProT α can stimulate the activated killer cell of lymphokine (lymphokien activated killer cells in early days in disease; LAK) activity; It is through increase lymphocyte and target cell combine so that increase that the secretion of IFN-γ and IL-2 realizes (20, Lopez-Rodriguez JL; Cordero OJ; Sarandeses C .Interleukin-2 killer cells:in vitro evaluation of combination with prothymosin alpha.Lymphokine Cytokine Res 1994; 13 (3): 175-182; 21, Baxevanis CN, Gritzapis AD, Dedoussis GV .Induction of lymphokine-activated killer activity in mice by prothymosin alpha.Cancer Immunol Immunother 1994; 38 (4): 281-286).If the secretion that can stimulate TNF-α and IL-2 when separately using ProT α when uniting use with IL-2, can increase the expression of NK cell CD25 and CD18/11 adhesion molecule.Under the condition that IFN-γ exists; ProT α is mainly through increasing the expression of CD54 (cell adhesion molecule part); Stimulate the combining of tumor cell of mononuclear cell and assembly, reach the removing tumor cell purpose (22, Baxevanis CN, Gritzapis AD; Spanakos G .Induction of tumor-specific T lymphocyte responses in vivo by prothymosin alpha.Cancer Immunol Immunother 1995; 40 (6): 410-418; 23, Lopez-Rodriguez JL; Cordero OJ; Sarandeses C .Interleukin-2 killer cells:in vitro evaluation of combination with prothymosin alpha.Lymphokine Cytokine Res 1994; 13 (3): 175-182; 24, Baxevanis CN, Gritzapis AD, Dedoussis GV .Induction of lymphokine-activated killer activity in mice by prothymosin alpha.Cancer Immunol Immunother1994; 38 (4): 281-286; 25, Voutsas IF; Baxevanis CN; Gritzapis AD; Et al.Synergy between interleukin-2 and prothymosin alpha for the increased generation of cytotoxic T lymphocytes against autologous human carcinomas.Cancer Immunol Immunother 2000,49 (8): 449-458).Recently research shows; ProT α in dna vaccination, can also play vaccine adjuvant effect (26, Jin Y; Cao C, Li P, Liu X; Huang W, Li C et al.Boosting immune response to hepatitis B DNA vaccine by coadministration of Prothymosin alpha-expressing plasmid.Clin Diagn Lab Immunol 2005; 12 (12): 1364-1369; 27, Shiau AL; Chen CC; Yo YT; Chu CY, Wang SY, Wu CL.Enhancement of humoral and cellular immune responses by an oral Salmonella choleraesuis vaccine expressing porcine prothymosin alpha.Vaccine 2005; 23 (48-49): 5563-5571.)。
But also have no at present about prophymosin-alpha and prevent the report relevant with treating hepatocarcinoma through the approach that suppresses the regulatory T lymphocyte activity.
Summary of the invention
First purpose of the present invention is to provide the purposes of prophymosin-alpha in suppressing the regulatory T lymphocyte activity.
Second purpose of the present invention is to provide the application of prophymosin-alpha in preparation prevention and treatment liver-cancer medicine.
Below provide the former method for preparing of recombined human prothymosin:
From normal Chinese's PBLC through after extracting RNA; Adopt the method for RT-PCR to obtain the peculiar prophymosin-alpha cDNA of Chinese; Utilize the technique for gene engineering vitro recombination to comprise the recombinant expression carrier of thymosin alpha protogene, expression vector is changed over to carry out the former albumen of clonal expression purification acquisition recombinant thymin alpha in the prokaryotic cell escherichia coli again.Detailed process is referring to Chinese patent (ZL2008.10072084.4; Inventor Zhou Kefu etc.)
Below provide the functional experiment of prophymosin-alpha in prevention of hcc and treatment:
1) preparation of H22 original position liver cancer mouse model: 15d behind the selection abdominal cavity inoculation H22; The mice with tumor that upgrowth situation is good, from the abdominal cavity with the aseptic absorption ascites of disposable syringe, with the normal saline diluting cells; Wash 2 times, reuse normal saline diluting cells number is 8 * 10
6Individual/ml, viable count is greater than 95%, and getting the 0.2ml tumor cell suspension, to be inoculated in the right axil of mice subcutaneous.After 0.3～0.4cm size is grown in inoculation Subcutaneous tumor on the 7th～8, separate tumor tissues, be cut into the tissue of 0.5mm size under the aseptic condition.
2) with the operation of beginning to speak under 30 mice aseptic conditions, the exposed portions serve liver advances liver to seal opening with biogum immediately tumor tissues with syringe needle, the abdominal cavity is sewed up again.
3) experiment is divided into groups: behind 30 orthotopic transplantation liver cancer mouse model mixings through operation, press A again, B, C, four groups every group 10 sub-cage rearings of D.A, the B group is the medication group, lumbar injection prophymosin-alpha dosage is respectively 150 μ g/ and 300 μ g/.C group is injected the normal saline of same dosage for matched group, and selecting 10 onesize mices in addition is the normal control group, not transplantation tumor but injecting normal saline equally.Surveyed body weight 1 time in per 3 days.Experiment was the cycle with 21 days.
4) index determining: experiment finishes back separation liver tumor weighs, and measures the ascites volume.
5) the lymphocytic detection of mouse spleen regulatory T:
(1) aseptic separation mouse spleen grinds the back with the sterile needle tube head and filters with 200,300 order filter membranes, and filtrate is removed erythrocyte and obtained lymphocyte solution under the ammonium chloride solution effect.
(2) separate the pure lymphocyte of acquisition with lymphocyte separation medium.
(3) the lymphocytic fluorescent antibody of applying detection regulatory T, CD4, CD25, CD127-FITC be after labelling carries out trichrome stain respectively, up flow type cell instrument analytic record result.Use homotype as negative control simultaneously.
Can adopt surgical method successfully to set up and transplant hepatocarcinoma H22 model in the body.
Adopt the lumbar injection prophymosin-alpha, observe the effect of this recombiant protein in prevention and treatment hepatocarcinoma, comprise the size of liver tumor and the formation that prevents ascites.
Through the injection for curing and the prevention of prophymosin-alpha, the growth that can obviously suppress liver tumor can reduce the formation of ascites simultaneously.
Through detecting the lymphocytic level of mouse spleen regulatory T, prove that the lymphocytic level of hepatocarcinoma matched group spleen regulatory T is obviously than the lymphocytic level height difference heteropole of normal mouse spleen regulatory T remarkable (P＜0.01); The lymphocytic level of medication group regulatory T is then obviously low than normal control group and experiment contrast group, and difference is (P＜0.01) extremely significantly.
Experimental result is found; The former growth that can obviously suppress rat liver cancer H22 in the body of recombinant thymin alpha; Through analyzing mouse spleen regulatory T lymphocyte level, the mechanism that further confirms this effect is through downward modulation mouse spleen regulatory T lymphocyte level, thereby removes the rejection condition to immune function of mice; Transfer the normal autoimmune function of body, reach the purpose that suppresses the interior tumor cell growth.
Interpretation of result proves, the recombined human prothymosin former in prevention and treatment body hepatocarcinoma have obvious effect, effect is remarkable.And clearly illustrate the wherein mechanism of action.Therefore, utilize this albumen to can be used in the generation and the development of prevention and treatment hepatocarcinoma.Significant for exploitation corresponding prevention and medicine.
This shows that prophymosin-alpha can be used for suppressing the regulatory T lymphocyte activity, and can in preparation prevention and treatment liver-cancer medicine, use.
Said prophymosin-alpha comprises the prophymosin-alpha of all mammals extractions and the prophymosin-alpha of gene engineering expression.
Said hepatocarcinoma comprises various hepatocarcinoma.
Said medicine comprises oral drugs, subcutaneous injection medicine and intraperitoneal injection of drugs etc.
Description of drawings
Fig. 1 suppresses the effect of rat liver cancer cell H22 (orthotopic transplantation type) in the ProT α body (K1～K3 represents the normal mouse liver; C1～C3 represents control group mice liver and growing tumors; T1～T3 medication group mouse liver and growing tumors).
Fig. 2 suppresses the effect analysis of rat liver cancer cell H22 (orthotopic transplantation type) in the ProT α body (K represents the normal mouse liver; C represents control group mice tumor gram number; T medication group mouse tumor gram number; * represent significant difference).
Fig. 3 is flow cytometry analysis figure, the lymphocyte of R1 representative delineation, and abscissa FS represents (forword scatter) forward scattering light; Vertical coordinate ss represents side scattered light (side scatter).
Fig. 4 is flow cytometry analysis figure, the cell that on behalf of CD4+/CD127, R2 express; On behalf of two of marked by fluorescein isothiocyanate, abscissa FITC (Fluorescein Isothiocyanate) resist; CD4: represent leukocyte differentiation antigen; Vertical coordinate PE-CD127: represent the painted fluorescence two of PE to resist.
Fig. 5 is flow cytometry analysis figure, the cell that on behalf of CD25, R3 express.On behalf of two of marked by fluorescein isothiocyanate, abscissa FITC (Fluorescein Isothiocyanate) resist; CD4: represent leukocyte differentiation antigen; Vertical coordinate PRP-CY5-CD25 represents leukocyte differentiation antigen
Fig. 6 is flow cytometry analysis figure, and homotype negative control, abscissa are represented FITC (Fluorescein Isothiocyanate), and two of marked by fluorescein isothiocyanate resists; CD4: represent leukocyte differentiation antigen.Vertical coordinate PRP-CY5-CD25 represents leukocyte differentiation antigen
Fig. 7 is flow cytometry analysis figure, normal mouse 1 regulatory T lymphocyte expression; On behalf of two of marked by fluorescein isothiocyanate, abscissa FITC (Fluorescein Isothiocyanate) resist; CD4: represent leukocyte differentiation antigen.Vertical coordinate PRP-CY5-CD25 represents leukocyte differentiation antigen.
Fig. 8 is flow cytometry analysis figure, normal mouse 2 regulatory T lymphocyte expressions; On behalf of two of marked by fluorescein isothiocyanate, abscissa FITC (Fluorescein Isothiocyanate) resist; CD4: represent leukocyte differentiation antigen.Vertical coordinate PRP-CY5-CD25 represents leukocyte differentiation antigen.
Fig. 9 is flow cytometry analysis figure, normal mouse 3 regulatory T lymphocyte expressions; On behalf of two of marked by fluorescein isothiocyanate, abscissa FITC (Fluorescein Isothiocyanate) resist; CD4: represent leukocyte differentiation antigen.Vertical coordinate PRP-CY5-CD25 represents leukocyte differentiation antigen.
Figure 10 is flow cytometry analysis figure, control group mice 1 regulatory T lymphocyte expression; On behalf of two of marked by fluorescein isothiocyanate, abscissa FITC (Fluorescein Isothiocyanate) resist; CD4: represent leukocyte differentiation antigen.Vertical coordinate PRP-CY5-CD25 represents leukocyte differentiation antigen.
Figure 11 is flow cytometry analysis figure, control group mice 2 regulatory T lymphocyte expressions; On behalf of two of marked by fluorescein isothiocyanate, abscissa FITC (Fluorescein Isothiocyanate) resist; CD4: represent leukocyte differentiation antigen.Vertical coordinate PRP-CY5-CD25 represents leukocyte differentiation antigen.
Figure 12 is flow cytometry analysis figure, control group mice 3 regulatory T lymphocyte expressions; On behalf of two of marked by fluorescein isothiocyanate, abscissa FITC (Fluorescein Isothiocyanate) resist; CD4: represent leukocyte differentiation antigen.Vertical coordinate PRP-CY5-CD25 represents leukocyte differentiation antigen.
Figure 13 is flow cytometry analysis figure, medication group mice 1 regulatory T lymphocyte expression; On behalf of two of marked by fluorescein isothiocyanate, abscissa FITC (Fluorescein Isothiocyanate) resist; CD4: represent leukocyte differentiation antigen.Vertical coordinate PRP-CY5-CD25 represents leukocyte differentiation antigen.
Figure 14 is flow cytometry analysis figure, medication group mice 2 regulatory T lymphocyte expressions; On behalf of two of marked by fluorescein isothiocyanate, abscissa FITC (Fluorescein Isothiocyanate) resist; CD4: represent leukocyte differentiation antigen.Vertical coordinate PRP-CY5-CD25 represents leukocyte differentiation antigen.
Figure 15 is flow cytometry analysis figure, medication group mice 3 regulatory T lymphocyte expressions; On behalf of two of marked by fluorescein isothiocyanate, abscissa FITC (Fluorescein Isothiocyanate) resist; CD4: represent leukocyte differentiation antigen.Vertical coordinate PRP-CY5-CD25 represents leukocyte differentiation antigen.
Figure 16 is that (K represents normal mouse, and C represents matched group, and prophymosin-alpha group mice is used in the T1 representative for medication group, matched group and normal mouse spleen regulatory T lymphocyte expression comparative analysis figure; Treg/R1 represents the ratio of regulatory T lymphocyte and all TLCs, and * represents significant difference; * represents utmost point significant difference; * * represents utmost point significant difference).
Find from the result of above accompanying drawing; The former growth that can obviously suppress rat liver cancer H22 in the body of recombinant thymin alpha; Confirm further that through analyzing mouse spleen regulatory T lymphocyte level the mechanism of this effect is through downward modulation mouse spleen regulatory T lymphocyte level; Thereby remove rejection condition, transfer the normal autoimmune function of body immune function of mice.Reach the purpose that suppresses the interior tumor cell growth.
Comprehensive above interpretation of result proof recombined human prothymosin is former in prevention with treat that hepatocarcinoma has obvious effect in the body, and effect is remarkable.And clearly illustrate the wherein mechanism of action.Therefore, utilize this albumen to can be used in the generation and the development of prevention and treatment hepatocarcinoma.Significant for exploitation corresponding prevention and medicine.
The specific embodiment
Following examples will combine accompanying drawing that the present invention is further described.
The materials and methods explanation that embodiment adopted as follows.
1. experiment is drawn materials
The former preparation of recombined human prothymosin: from normal Chinese's PBLC through after extracting RNA; Adopt the method for RT-PCR to obtain the peculiar prophymosin-alpha cDNA of Chinese; Utilize the technique for gene engineering vitro recombination to comprise the recombinant expression carrier of thymosin alpha protogene, expression vector is changed over to carry out the former albumen of clonal expression purification acquisition recombinant thymin alpha in the prokaryotic cell escherichia coli again.Detailed process can be referring to Chinese patent (ZL2008.10072084.4; Inventor Zhou Kefu etc.).
Experimental animal is a Kunming mouse, the SPF level, and (20 ± 2) g, in age in 6-7 week, male, Xiamen University's Experimental Animal Center provides, the quality certification number 0501452.Tumor kind Mus is provided by Xiamen University's medical college Experimental Animal Center.
CD4 FITC (rat anti-mouse); CD25 PERCP-CY5.5 (rat anti-mouse); CD127 PE (rat anti-mouse); Homotype IGG2A FITC Control fluorescence monoclonal antibody is all available from BD company.
2. implementation method
2.1 tumor control rate experiment
The former albumen of recombined human prothymosin suppresses rat liver cancer H22 and suppresses the experiment of regulatory T lymphocyte activity; 15d behind the selection abdominal cavity inoculation H22; The mice with tumor that upgrowth situation is good, from the abdominal cavity with the aseptic absorption ascites of disposable syringe, with the normal saline diluting cells; Wash 2 times, reuse normal saline diluting cells number is 8 * 10
6Individual/ml, viable count is greater than 95%, and getting the 0.2ml tumor cell suspension, to be inoculated in the right axil of mice subcutaneous.After 0.3～0.4cm size is grown in inoculation Subcutaneous tumor on the 7th～8; Separate tumor tissues; Be cut into the tissue of 0.5mm size under the aseptic condition, simultaneously with the operation of beginning to speak under 30 mice aseptic conditions, exposed portions serve liver; Advance liver to seal opening with biogum immediately tumor tissues with syringe needle, again the abdominal cavity is sewed up.Behind 30 orthotopic transplantation liver cancer mouse model mixings, press A again, B, C, 4 groups every group 10 sub-cage rearings of D through operation.A, the B group is the medication group, lumbar injection prothymosin dosage is respectively 150 μ g/ and 300 μ g/.C group is injected the normal saline of same dosage for matched group, and selecting 10 onesize mices in addition is the normal control group, not transplantation tumor but injecting normal saline equally.Surveyed body weight 1 time in per 3 days.Experiment was the cycle with 21 days.Dissecting mice after experiment finishes separates liver neoplasm and surveys tumor weight.
Statistical analysis adopts Graph prime statistical software to analyze, and significant difference is by (P＜0.05); Difference is extremely significantly by (P＜0.01).
The result sees Fig. 1 and Fig. 2, and as can be seen from the figure medication group tumor weight is obviously little than matched group, significant difference (P＜0.05)
2.2 prothymosin is to the lymphocytic influence of mouse spleen regulatory T
Above experiment is separated the mouse spleen that obtains, behind grinding and 200 order nylon wires and 300 order nylon net filters, adopt the mouse lymphocyte separating medium to separate and obtain spleen lymphocyte, with in the PBS liquid of 2% paraformaldehyde (4%).By carrying out adopting flow cytometer to carry out trichrome stain after the fluorescent antibody effect, detect the normal mouse spleen respectively, medication group and the lymphocytic ratio of matched group spleen regulatory T again.
Represent the lymphocytic flow cytometry result of regulatory T to see Fig. 3 (for lymphocyte), Fig. 4 (cell that CD4+/CD127 expresses), Fig. 5 (cell that CD25 expresses);
Adopt the homotype negative control figure of no antibody to see Fig. 6
See Fig. 7～9 with the isolating regulatory T lymphocyte of normal mouse spleen, can find out that normal mouse regulatory T lymphocyte all can detect;
Transplanted hepatoma mouse spleen regulatory T lymphocyte is seen Figure 10～12, finds out that from figure the lymphocytic expression of hepatocarcinoma matched group regulatory T is obviously high than normal group, and difference is (P＜0.01) extremely significantly.
Same transplanted hepatoma mice, but intervene with prophymosin-alpha, the lymphocytic level of the regulatory T in its spleen is seen Figure 13～15, from figure, finds out that its level is obviously low than hepatocarcinoma matched group.Difference is (P＜0.01) extremely significantly
The three groups of lymphocytic comparative analysis as a result of mice regulatory T figure see Figure 16, from figure, see that normal mouse regulatory T lymphocyte level is moderate, and matched group is the highest, and the medication group is minimum.There are utmost point significant difference (P＜0.01) in medication group and matched group;
Above result proves that first prophymosin-alpha is through suppressing the effect that the lymphocytic level of regulatory T reaches the growth that suppresses hepatocarcinoma.
Claims (6)
1. the purposes of prophymosin-alpha in suppressing the regulatory T lymphocyte activity.
2. purposes as claimed in claim 1 is characterized in that said prophymosin-alpha comprises the prophymosin-alpha of all mammals extractions and the prophymosin-alpha of gene engineering expression.
3. the application of prophymosin-alpha in preparation prevention and treatment liver-cancer medicine.
4. application as claimed in claim 3 is characterized in that said prophymosin-alpha comprises the prophymosin-alpha of all mammals extractions and the prophymosin-alpha of gene engineering expression.
5. application as claimed in claim 3 is characterized in that said hepatocarcinoma comprises various hepatocarcinoma.
6. application as claimed in claim 3 is characterized in that said medicine comprises oral drugs, subcutaneous injection medicine and intraperitoneal injection of drugs.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1076361A (en) * | 1993-01-19 | 1993-09-22 | 祁岩超 | The preparation method of human placental thymosin |
| CN1094411A (en) * | 1993-02-03 | 1994-11-02 | 阿尔法-1生物医药品有限公司 | Thymosin alpha-1 derivatives |
-
2012
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1076361A (en) * | 1993-01-19 | 1993-09-22 | 祁岩超 | The preparation method of human placental thymosin |
| CN1094411A (en) * | 1993-02-03 | 1994-11-02 | 阿尔法-1生物医药品有限公司 | Thymosin alpha-1 derivatives |
Non-Patent Citations (2)
| Title |
|---|
| 《中国优秀博硕士学位论文全文数据库（硕士）基础科学辑》 20021105 邱磊 胸腺素alpha原的生物学活性及DNA芯片分析研究 第50-55页讨论部分 3-6 第2002年卷, 第01期 * |
| 邱磊: "胸腺素α原的生物学活性及DNA芯片分析研究", 《中国优秀博硕士学位论文全文数据库（硕士）基础科学辑》 * |
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Application publication date: 20120718 |