| BACKGROUND&OBJECTIVEThe colon cancer (Colorectal cancer, CRC) is one of most common digestive tract malignant tumors. In recent years, as a result of the life food habit change’s reason, our country colon cancer’s disease incidence rate and the mortality rate assumed gradually the trend of escalation unceasingly, but made a sneak attack and the shift causes the primary cause which the patient died. The tumor occurrence and the development are more than factors, the multi-steps, the polygenes combined action complex process. Between the tumour cell and host’s interaction is the tumor has the development intrinsic factor, the tumour cell shift has certain organ tendentiousness, this is possibly the result which the lump cell and the specific organ micro environment interaction chooses mutually. The tumor shift is decided by tumour cell’s molecular characteristic and its secretion factor to the periphery intersititial cell’s influence, including the blood vessel, the knot contracts the organization and the immune cell and so on. The special tumor micro environment affects tumour cell’s growth through each kind of adjustment mechanism to develop and to make a sneak attack shifts. Therefore, not only the tumor shift tumour cell’s biology characteristic and the hereditary property decided that fundamentally speaking is the cancer cell and the host interaction result, the host even plays the decisive effectThe hematopoietic ancestor cell (hematopoietic progenitor cell) is the Hemopoietic tissue has the self-duplication and the split up potential initial cell. The initial CD34+antigen was considered that is the hematopoietic ancestor cell’s important symbol, this antigen is exuberant in the primitive hematopoietic cell expression, latter weakens gradually along with the cell differentiation. To the ancestor cell surface symbolic antigen’s preliminary study discovery, the CD133+cell compares the CD34+cell to have the stronger multiplication potential, and contains has the higher proportion long-term raise outset cell, the CD133+cell possibly had under the normal marrow micro environmental condition compared to the CD34+cell stronger hastens and the migration ability, the CD133+hemopoiesis ancestor cell has possibly represented the more primitive hematopoietic cell.Kaplan RN had discovered the blood vessel bast ancestor cell which the VEGFR1masculine hemopoiesis ancestor cell (vascular endothelial growth factor receptor1positive hematopoietic progenitor cell, VEGFR1+HPC) plays the decision in the animal tumor shift the role, VEGFR1+HPC discovered before by no means (endothelial progenitor cell, EPC) is the VEGFR2masculine hemopoiesis ancestor cell, but simultaneously has CD133and the VEGFR1two kind of mark one kind of hematopoietic ancestor cells, because the abdominal cavity injects the VEGFR2immune body, cannot block the VEGFR1+HPC cell bunch to form and the lump shift stove formation, can only limit the tumor shift stove growth. Using Beta half young glycosidase mark marrow cell transplant’s mouse Lewis lung cancer model, the mouse B16melanin lump model and the c-myc transgene mouse spontaneous lymphoma model discovered:VEGFR1+HPC first in the specific organ formation cell bunch (VEGFR1+HPC cellular clusters), prepared for the lump cell easy to form the shift the micro environment, these cell bunch formed the spot to be consistent with the tumor shift common spot. The tumor shifts to some kind of specific organ’s cell and the molecular mechanism is not very clear, but many tumors have shift to some specific organ’s tendency. The dependence tumor shift related cell label-VEGFR1+hemopoiesis ancestor cell forms the shift in the target organ the micro environment, promoted the tumor attack and the shift. But after the present research uses the animal the lethal radiation, carries on research technique which the marrow transplants, after various organs existence radiation damage, possibly will create the non-specificity marrow mobilization and the marrow originates cell’s accumulation, and not yet will test the level at the person cancer to obtain the confirmation. Person CD133high compatible peptide TR-7is this topic-based group earlier period in screens the mouse high compatible peptide in the foundation to screen, after the confirmation can with the person cell’s on CD133special union.CONTENT1. The CD133+hemopoiesis ancestor cell multiplies, the attack to the colon cancer and the shift influence, expounded that the CD133+hemopoiesis ancestor cell’s mobilizes the CD133+hemopoiesis ancestor cell in colon cancer shift main biology function as well as the colon cancer to enter the blood circulation then to enter shifts stove’s principle, lays the foundation for the revelation colon cancer shift mechanism and the anti-shift treatment2. Person CD133the high compatible peptide TR-7in vitro biology effect the way which altogether raises using the CD133high compatible peptide and the person tumour cell separately to its multiplication, the adherency and in vitro motor ability carries on the examination Method1. CD133+hemopoiesis ancestor cell’s screening, appraisal and raise is from the South hospital obstetrics newborn embryo fresh navel blood specimen20examples, the antifreeze preservation, and in the navel blood gathers in the latter2hours, uses the person lymphocyte parting method, the separation takes out the single nuclear cell, using the CD133+immunity magnetism bead parting method, separates the CD133+hemopoiesis ancestor cell again, and gives the class type cell meter and the immune cell dyeing separately analyzes and appraises in the sample size the CD133+cell content, simultaneously during the reduction induces the CD133+differentiation, raises the CD133+hemopoiesis ancestor cell.2. CD133+hemopoiesis ancestor cell to colon cancer cytobiology characteristic influence Altogether hasten the raise the CD133+hemopoiesis ancestor cell and SW480, and adheres ability with the MTT law examination tumour cell’s multiplication the influence, with the Transwell cab law examination tumour cell’s migration ability’s change, understands VEGFR-1, MMP-2, the E-Cadherin expression situation with Western blot. Observes around the CD133+hemopoiesis ancestor cell function the cell multiplication, in vitro attack to change the3. CD133high compatible peptide to oppress to the CD133+hemopoiesis ancestor cell function anti-, in the previous step experiment, joins the CD133high compatible peptide to form the experiment using the plate clone to examine the CD133high compatible union peptide to the large intestine cancer cell in vitro multiplication ability influence; Determines the cell using the cell adherency experiment in the textile fiber adhesion protein (Fibronectin, FN) on coherence, the application movement cab experiment, the cell scratch experiment examines this short peptide to the large intestine cancer cell movement migration ability influence. Examination forms the experiment, the cell adherency experiment, the cell movement experiment’s data using One-Way the ANOVA to the plate clone to carry on processing and the analysis, using LSD, Dunnett’s T3multiple compares further comparative analysisRESULTS1. CD133+hemopoiesis ancestor cell’s screening, appraisal and raisefrom the South hospital obstetrics, navel blood specimen altogether20(18success extraction fresh cell,2defeats),50-80ml/shares,5%citric acid antifreeze, the fresh navel blood gathers in latter2h, separates the CD133+hemopoiesis ancestor cell. Single nuclear cell number average which obtains from the fresh navel blood for (1.43±0.17) x107/ml, results in CD133+after the CD133magnetism bead separation system separation to concentrate cell’s mean value for (1.32±0.01) x105/ml, in the fresh navel blood single nuclear cell’s CD133+cell accounts for the proportion for (0.8±0.01)%, appraises the CD133+CD34+cell purity after the class type cell meter to achieve above80%. CD133+cell culture one week later, the immunocytochemistry dyes CD133, under the microscope observes the CD133masculine cell to count>90% 2. the CD133+hemopoiesis ancestor cell can obviously promote the SW480cell’s growth to the colon cancer cytobiology characteristic influenceCD133+hemopoiesis ancestor cell (n=60, Z=-6.106, P=0.000), can promote tumour cell’s morphology change, namely tumour cell’s nuclear engrain, the heterogeneous type is obvious, the cell assumes the polygon or the anomalous shape, can obviously promote SW480cell’s migration ability; The cell adheres the experiment to indicate that includes the CD133+hemopoiesis ancestor cell’s experimental group its tumour cell to adhere ability to be higher than the control group obviously (n=60, Z=-3.679, P=0.000).(I.e. concentration protein) examines MMP-2, VEGFR-1and the E-Cadherin protein expression situation using the acetone precipitation method extraction cell total protein discovered that in the experimental group3kind of proteins express the water to be higher than the control group equally.3. the different density’s TR-7short peptide to each group of colon cancer HT-29cell multiplication ability which altogether raises does not have the obvious function, the multiplication ability difference not to have statistics significance (F=1.473, P=0.347). Moreover each group of tumour cell’s clone speed of formation experimental result indicated that four group of cell multiplication ability the difference has statistics significance (F=0.174, P=0.000). Through to adhesive capacity’s examination, different density TR-7to SW480, the SW620cell adhesive capacity influence’s difference does not have statistics significance (Fht-29=0.428, P ht-29=0.738; Fsw620=3.109, P sw620=0.089). The Transwell cell movement experiment demonstrated after different concentration gradient TR-7altogether raises SW480, SW620, the Lovo cell, passes through the membrane the cell number obviously to reduce, the different density group has the remarkable difference, the difference has statistics significance. And the TR-7 density is bigger puts on the membrane the cell number to be less, is more obvious to the cell movement ability’s influence. The cell scratch experiment further confirmed that along with TR-7short peptide density’s increase, the cell number which the lovo cell scratch spot migrates are less.CONCLUSION1.1, CD133+hemopoiesis ancestor cell may obviously promote the person colon cancer cell’s multiplication and attack ability, the CD133+hemopoiesis ancestor cell and the person colon cancer cell bare mouse altogether transplants has presses the shift function, proved initially, the person hemopoiesis ancestor cell possibly forms to the person colon cancer cell shift has the material effect2. the CD133+hemopoiesis ancestor cell may with the colon cancer cell direct action, be able to promote the shift related gene and blood vessel production factor MMP-2, the VEGFR-1protein high expression, simultaneously adjusts E-Cadherin the protein expression, indicated initially:The CD133+hemopoiesis ancestor cell presses the shift function to be possible and shift multiple coefficient of correlation MMP-2, VEGFR-1and the E-Cadherin expression concerns.3. CD133high compatible peptide CR-7can block the CD133hemopoiesis ancestor cell to tumour cell’s promoter action... |
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