CN103751221B - The application in the medicine of preparation treatment liver cancer of the SIRT1 inhibitor associating Sorafenib - Google Patents
The application in the medicine of preparation treatment liver cancer of the SIRT1 inhibitor associating Sorafenib Download PDFInfo
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Abstract
The invention belongs to chemical field, it is specifically related to the application in the medicine of preparation treatment liver cancer of the SIRT1 inhibitor associating Sorafenib, the method utilizing the present invention can suppress the propagation of liver-cancer stem cell, differentiation and Clone formation, improve liver-cancer stem cell and to the sensitiveness of Sorafenib and reduce the liver-cancer stem cell drug resistance to Sorafenib, the Sorafenib damage to liver-cancer stem cell can also be strengthened simultaneously, from source, kill tumor stem cell, bring new hope for thorough tumor eradication.
Description
Technical field
The invention belongs to chemical field, particularly to the application in the medicine of preparation treatment liver cancer of the SIRT1 inhibitor associating Sorafenib.
Background technology
Liver cancer is the malignant tumour in the 5th, the ranking world, main distribution Asia and country in Southeast Asia, M & M remains high, and the traditional remedies such as operation, radiotherapy, chemotherapy is difficult to thoroughly eradicate, treatment to liver cancer lacks effective medicine, and tracing it to its cause is that medicine easily produces inherent drug resistance.
The pathogenesis of liver cancer has complexity, the transfer that development and cancer cell occur of its cancer cell is closely-related with intracellular various Cell signal propagation pathways and growth factor etc., wherein owing to there is multiple key gene or signaling molecule in Cell signal propagation pathways, such as Ras/Raf/MAPK, PI3K/AKT/mTOR, the signal paths such as WNT/ β-catenin, Hedgehog[58,59].Wherein, Ras/Raf/MAPK, PI3K/AKT/mTOR signal path in the pathogenesis of liver cancer in occupation of key player.Ras/Raf/MAPK signal path can activate in liver cancer the growth factor of upstream also can activated tumor suppressor, and PI3K/AKT/mTOR signal transduction pathway also plays an important role in liver cancer, it is possible to breed and activate the HCC of 30-50%[60].Little molecular targeted agents is how U.S. lucky, also known as Sorafenib (sorafenib), the propagation of tumour cell can be suppressed, it is a kind of oral Mutiple Targets multi-kinase inhibitor, can be with targeting growth factor on the serine/threonine kinase of tumour cell and receptor tyrosine kinase and tumor vessel.Research report both at home and abroad, Sorafenib can delay the progress of liver cancer and can extend patients with terminal life cycle.At present, Sorafenib is except, in addition to the treatment of liver cancer patient, also using in breast cancer, Patients with Gastric Cancer.But cancer patient easily produces drug dependence when using Sorafenib, it has been investigated that tumour cell contains the tumor stem cell of trace, this kind of cell has tolerance, the feature such as insensitive to conventional radiotheraphy, chemotherapy, is the root of tumour drug dependence, Preventive.Therefore, it is badly in need of researching and developing and from source, the targeted drug of tumor stem cell can being killed tumor stem cell, new hope will be brought for thorough tumor eradication.
Summary of the invention
In view of this, it is an object of the invention to provide the application in the medicine of preparation treatment liver cancer of the SIRT1 inhibitor associating Sorafenib, being bred by suppression tumor stem cell, broken up, and kill tumor stem cell from source, final acquisition reaches to treat the purpose of liver cancer.
For achieving the above object, the present invention provides following technical scheme:
The application in the medicine of preparation treatment liver cancer of the SIRT1 inhibitor associating Sorafenib.
Preferably, SIRT1 inhibitor associating Sorafenib application in the medicine of preparation suppression liver-cancer stem cell self.
It is furthermore preferred that described liver-cancer stem cell self is the propagation of liver-cancer stem cell, differentiation and Clone formation.
Preferably, described SIRT1 inhibitor is Tenovin-6 or the slow virus of interference SIRT1 expression, it is also possible to for other SIRT1 inhibitor.
Preferably, the slow virus that described interference SIRT1 expresses contains Lv-sh-SIRT1 plasmid.
It is furthermore preferred that the slow virus that described interference SIRT1 expresses is obtained through packaging, purifying by after Lv-sh-SIRT1 plasmid transfection 293T cell by Lv-sh-SIRT1 plasmid.
The beneficial effects of the present invention is: the invention discloses the application in the medicine of preparation treatment liver cancer of the SIRT1 inhibitor associating Sorafenib, utilize the interference of SIRT1 inhibitor or propagation, differentiation and the Clone formation of suppression SIRT1 expression inhibiting liver-cancer stem cell, improve liver-cancer stem cell simultaneously and to Sorafenib sensitiveness and strengthen the Sorafenib damage to liver-cancer stem cell, the method utilizing the present invention can also reduce the liver-cancer stem cell drug resistance to Sorafenib, kill liver-cancer stem cell, from source, kill tumor stem cell, bring new hope for thorough tumor eradication.
Accompanying drawing explanation
In order to make the purpose of the present invention, technical scheme and beneficial effect clearer, the present invention provides drawings described below:
Fig. 1 is to detect the expression of SIRT1 in clinical hepatic tissue sample (A: SABC detection SIRT1 is in hepatic carcinoma tissue and the expression and localization of cancer beside organism；The statistics that B:SIRT1 expresses at hepatic carcinoma tissue and cancer beside organism；The expression of C:Kaplan-Meier methods analyst SIRT1 and the relation of liver cancer patient prognosis).
Fig. 2 is that to the drug resistance of Sorafenib, (A: after disturbing the expression of SIRT1 in liver-cancer stem cell, detection Sorafenib processes the impact on cell clonal formation power to liver-cancer stem cell；B: after suppressing the expression of SIRT1 in liver-cancer stem cell, detection Sorafenib processes the impact on cell clonal formation power；After C: Immunofluorescence test interference SIRT1 expresses, detection Sorafenib processes the impact on cellular damage).
Fig. 3 is that the associating Sorafenib of expressing suppressing SIRT1 in liver-cancer stem cell processes the impact on its drug resistance of cell.
Fig. 4 is that the associating Sorafenib of expressing disturbing SIRT1 in liver-cancer stem cell processes the transplantable tumor impact on transplantable tumor weight or volume.
Fig. 5 is that the associating Sorafenib of expressing disturbing SIRT1 in liver-cancer stem cell processes the impact damaging transplantable tumor form and transplantable tumor.
Fig. 6 is expression (A:Kaplan-Meier methods analyst liver cancer patient postoperative SIRT1 expression and the relation of life span of the tissue specimen detection SIRT1 of liver cancer patient associating Sorafenib treatment；After the associating Sorafenib treatment of B:Kaplan-Meier methods analyst liver cancer patient, SIRT1 expresses the relation with life span).
Detailed description of the invention
Below in conjunction with accompanying drawing, the preferred embodiments of the present invention are described in detail.The experimental technique of unreceipted actual conditions in embodiment, generally according to normal condition, the such as condition described in Molecular Cloning: A Laboratory guide (third edition, J. Pehanorm Brooker etc. writes), or according to the condition proposed by manufacturer.
The liver-cancer stem cell model liver-cancer stem cell T1224 that the present invention uses is built by this laboratory, this model contains the Lentiviral (Nanog/GFP reporting system) that Nanog promoter regulation GFP reporter gene builds, its construction method sees Shan et al.2012, hepatology, 56:1014-1014；Lv-sh-Scramble plasmid and Lv-sh-SIRT1 plasmid are built by this laboratory, and its building process is specifically shown in Shan et al.2012, hepatology, 56:1014-1014；HEK293 cell is preserved by this laboratory.
The Envision that the present invention usesTMWith DAB nitrite ion purchased from DAKO company of Denmark；EDTA(pH8) antigen retrieval buffers steps neoformation technology development co. purchased from Foochow.Sorafenib (sorafenib) tablet is purchased from Germany Bayer Schering pharma AG company；TV-6(Tenovin6) purchased from Santa Cruz company of the U.S.；Mouse-anti people's γ-H2AX antibody is purchased from Millipore company of the U.S..
One, the expression of SIRT1 is on liver cancer patient clinical pathological characteristic and the impact of patient's prognosis
Existing research shows, SIRT1 can be acted on by the activation plays " oncogene " of regulation p53, the function of " tumor suppressor gene " can be played by suppressing the expression of some tumour oncogenes again, SIRT1 be on earth " oncogene " also or " tumor suppressor gene ", the always focus of research.Therefore, the present invention is by collecting southwest hospital of Third Military Medical University in January, 2000 to the hepatocellular carcinoma paraffin specimen cancerous tissue of in December, 2005 surgery excision and each 148 examples of liver cancer cancer beside organism that match, and patient is followed up a case by regular visits to 5-10, all samples are after southwest hospital hepatobiliary surgery, all samples are fixed through 10% formalin solution, FFPE, 4～5 μm serial section, then utilize immunohistochemical method detection SIRT1 at the expression of 148 example liver cancer patients, analyze SIRT1 expression and clinical prognosis again, the correlation of pathological hallmarks.Immunohistochemical method particularly includes: cancerous tissue and cancer beside organism's organization chip instrument (aperture 0.6 μm) of 148 example liver cancer patients being taked cancerous tissue and other each two points of cancer, observes after H&E dyes, result is as shown in A in Fig. 1.
Immunohistochemistry results interpretation: SIRT1 stained positive signal framing is in nucleus, the palm fibre do not waited in the depth, yellow particle.Using sxemiquantitative point system to mark coloration result, every section is all observed 3 times at different time, remembers that mean value, score are divided two parts to form by cell dyeing percentage and staining power:
Cell dyeing percentage score:<5% cell positive is 0 point, and 5%～25% cell positive is 1 point, and 26%～50% cell positive is 2 points, and 51%～75% cell positive is 3 points,>75% cell positive is 4 points；
Staining intensity of cells is scored: faint yellow is 1 point, and brown color is 2 points, and brownish black is 3 points；
Two split-phases are taken advantage of as total score: 0～1 is divided into negative (-), 2～4 be divided into the weak positive (+), 5～8 are divided into positives (++), and 9～12 are divided into strong positive (+++).
All calculating data all represent with means standard deviation (x ± s), SPSS16.0 Mathematical Statistics Analysis software is used to carry out independent samples t test (between two groups, average ratio is relatively), P < 0.05 is that significant difference is notable, and P < 0.01 and P < 0.001 is significant difference highly significant.
Statistics shows, SIRT1 is at 67%(99/148) cancerous tissue in express, and the positive rate in cancer side is 31%(48/148), and SIRT1 is at the positive expression rate difference highly significant (B in Fig. 1) (p < 0.0001) of cancerous tissue Yu cancer beside organism, use the expression of Kaplan-Meier methods analyst SIRT1 and the relation of liver cancer patient Postoperative determination, as shown in C in Fig. 1, the survival of patients time of cancerous tissue SIRT1 expression feminine gender is considerably longer than SIRT1 and expresses positive patient (p < 0.031).Express and clinicopathologia correlation by analyzing SIRT1, the results are shown in Table 1.
The table 1 expression at liver cancer patient SIRT1 and the correlation of clinicopathological parameters
Note:(1)*p<0.05,**P < 0.01 significant difference. (2) x2test.
As shown in Table 1, SIRT1 and histological grade, Capsular infiltration, blood vessel cancer embolus, there is the difference (p=0.003, p=0.015, p=0.005) of conspicuousness.This may be relevant in the effect of the generation of liver cancer, the performance of development with SIRT1.
Two, the expression of the SIRT1 research to liver cancer clinical treatment action target spot
The configuration of Sorafenib (sorafenib) solution: first Sorafenib tablet is milled into powder, is then configured to the storage liquid of 100mg/mL with the DMSO that volume fraction is 1%, is configured to required concentration with complete medium during use.
γ-H2AX antibody: γ-H2AX antibody dilutes for 1:500 by volume.
(1) packaging of slow virus Lv-sh-SIRT1 and Lv-sh-Scramble
Culture dish middle berth 2～5 × 10 at 10cm6Individual 293T cell (preferably 20 instead of before, the state of cell is required good, the speed of growth is very fast, otherwise the biggest on viral yield impact), the nutrient solution DMEM(v/v containing 10%FBS), when cell confluency to 70%, renew the fresh DMEM(v/v containing 10%FBS) nutrient solution, it is used for after 3-4h transfecting.
Take Lv-sh-SIRT1 or the Lv-sh-Scramble slow virus expression plasmid of 15-20 μ g, in mixed liquor, drip the CaCl of 50 μ L2.5M respectively2It is separately added into sterilized water again to cumulative volume 500 μ L, 500 μ L2 × HBS buffer solution (2 × HBS buffer solution shakes before using on turbula shaker) is added after mixing, it is subsequently placed on turbula shaker concussion more than 10s, after completion of dropwise addition, continues concussion, after room temperature is placed 5 minutes, liquid is added in the 293T cultivated, after adding, all around rocks mixing, the DMEM(v/v renewing fresh 10%FBS in after addition 6-12 hour) nutrient solution.
(2) collection of slow virus Lv-sh-SIRT1 and Lv-sh-Scramble and purifying
Start latter 48 hours, 72 hours respectively to collect a cell training liquid from transfection, after 48 hours collect, it is supplemented with the training liquid of 15mL, the virus collected centrifugal 10min under the conditions of 1000rpm is removed cell fragment, collect supernatant, and with the membrane filtration of 0.45 μm, filtrate is transferred to the evaporating column of Millopore, the liquid volume being centrifuged to the groove of upper strata at 5000rpm is concentrated into 1mL(and can add several times).Concentrate is dispensed, is placed in-80 DEG C of low temperature refrigerators and preserves.
(3) HCC T1224 selected by flow cytometry apoptosis Nanog masculine liver cancer stem cell (is designated as NanogPos), then by the Nanog of sortingPosIt is planted in 6 orifice plates, treats that cell length is to 1 × 106Time, by counting by cell seeding in 24 holes, every porocyte is 200, the repetition of 3, each sample, next day adds the Sorafenib of 0 μM and 2.5 μMs of concentration respectively, every 3d sucking-off nutrient solution, then add identical concentration, continuous processing 4 times, fix with the paraformaldehyde that mass fraction is 4% after i.e. 12 days, violet staining, observes and counts the cloning efficiency of every porocyte, and result is as shown in A and B in Fig. 2.From A in Fig. 2, concentration is that liver-cancer stem cell clonality is not affected by the Sorafenib of 2.5 μMs, and difference is the most notable, it may be possible to after Long-Time Service Sorafenib, liver-cancer stem cell creates drug resistance to Sorafenib.
Nanog according to method sorting same as described abovePos, then it is planted in 6 orifice plates, treats that cell length is to 1 × 106Time, add slow virus Lv-sh-SIRT1 particle, after 72 hours, streaming counting is planted in 24 holes, and every porocyte is 200, the repetition of 3, each sample.Next day adds the Sorafenib of 0 μM and 2.5 μMs of concentration respectively, every 3d sucking-off nutrient solution, then adds identical concentration, continuous processing 4 times, fixes with the paraformaldehyde that mass fraction is 4% after i.e. 12 days, violet staining, observing and count the cloning efficiency of every porocyte, result is as shown in A in Fig. 2.From A in Fig. 2, NanogPosAfter interference SIRT1 expresses, after adding 2.0 μMs of Sorafenibs process 12 days, the Clone formation power of liver-cancer stem cell is substantially less than the liver-cancer stem cell that interference SIRT1 expresses.Therefore, the expression reducing SIRT1 can reduce the liver-cancer stem cell tolerance to Sorafenib.
According to method sorting Nanog same as described abovePos, then it is planted in 6 orifice plates, treats that cell length is to 1 × 106Time, by counting by cell seeding in 24 holes, every porocyte is 200, the repetition of 3, each sample, adds the TV-6 that concentration is 2 μMs next day and processes 48 hours, then fix with the paraformaldehyde that mass fraction is 4%, violet staining, observes and counts the cloning efficiency of every porocyte.Then according to identical method, after processing cell 48 hours with the inhibitor TV-6 of the SIRT1 that concentration is 2 μMs, adding concentration is that 2.5 μMs of Sorafenibs process 12 days, then observes cloning efficiency, and its result is as shown in B in Fig. 2.
From B in Fig. 2, processing with the Sorafenib of 2.5 μMs for a long time and not affect liver-cancer stem cell Clone formation, after processing with the TV-6 that concentration is 2 μMs, liver-cancer stem cell clonality significantly reduces.And after disturbing SIRT1 to express, after adding 2.5 μMs of Sorafenibs process 12 days, the Clone formation of liver-cancer stem cell does not substantially less than interfere with the control group (p < 0.001) that SIRT1 expresses.Processing cell with the inhibitor TV-6 of SIRT1, add 2.5 μMs of Sorafenibs, after 12 days, its liver-cancer stem cell Clone formation is substantially less than the control group (p < 0.001) processed without TV-6 equally.Result shows, the high expressed of SIRT1 is the root that cancer treatment drug-Sorafenib treatment liver cancer produces resistance.
According to above-mentioned identical method, process Nanog with the Sorafenib of 0 μM, 1 μM, 5 μMs and 10 μMs respectivelyPosNanog after interference SIRT1 expressionPos, after 12 days, use EnvisionTMThe expression of immunohistochemistry technique detection DNA damage index γ-H2AX, after immunofluorescence dyeing, through confocal laser scanning microscope, result is as shown in C in Fig. 2.From C in Fig. 2, along with the increase of Sorafenib concentration, the damage of liver-cancer stem cell dramatically increases, and after interference SIRT1 expresses, the damage of liver-cancer stem cell is significantly higher than and does not interferes with the liver-cancer stem cell that SIRT1 expresses.Result shows, the expression of suppression SIRT1, can increase the sensitiveness treated liver-cancer stem cell to Sorafenib.
The Sorafenib sensitiveness to chemotherapeutics can be combined for further clear and definite SIRT1, thus provide effective clinical protocol for treatment liver cancer, by the Nanog of sortingPosCell is with doing following process respectively, and 2.0 μMs of TV-6 process 15 days；2.5 μMs of Sorafenibs process 15 days；After first processing 9 days with 2.5 μMs of Sorafenibs, then process 6 days with 2.0 μMs of TV-6；First process 9 days with 2.0 μMs of TV-6, then process 6 days with 2.5 μMs of Sorafenibs；By 2.0 μMs of TV-6 and 2.5 μMs of Sorafenib Combined Treatment, then statistics Clone formation situation, result is as shown in Figure 3.From the figure 3, it may be seen that it is less on the impact of its Clone formation individually to process liver-cancer stem cell with 2.5 μMs of Sorafenibs, and individually process, with 2.0 μMs of TV-6, the liver-cancer stem cell that the impact of its Clone formation is processed by liver-cancer stem cell more than 2.5 μMs of Sorafenibs；Show by the result of 2.0 μMs of TV-6 and 2.5 μMs of Sorafenib process liver-cancer stem cells simultaneously, first process cell 9 days with 2.5 μMs of Sorafenibs, the cell processed with 2.5 μMs of TV-6 again 6 days, less on the impact of liver-cancer stem cell Clone formation, and first process cell 9 days with 2.0 μMs of TV-6, the cell processed with 2.5 μMs of Sorafenibs again 6 days, the Clone formation of liver-cancer stem cell is affected bigger, but by 2.0 μMs of TV-6 and 2.5 μMs of Sorafenib Combined Treatment, the Clone formation of liver-cancer stem cell is affected maximum.Result above shows, the suppression expression of SIRT1 is also combined Sorafenib and can be strengthened the sensitiveness to chemotherapy of hepatocellular carcinoma medicine, brings hope for eradicating liver-cancer stem cell.
Three, SIRT1 is disturbed to combine the Sorafenib impact on subcutaneous transplantation knurl in immunodeficient mouse body
Showing according to foregoing study results, interference SIRT1 expresses or SIRT1 inhibitor is combined Sorafenib and can be significantly reduced the clonality of liver-cancer stem cell.For verifying above-mentioned conclusion further, in NOD/SCID Mice Body, verify the clonality that can significantly reduce liver-cancer stem cell in interference SIRT1 expression or SIRT1 inhibitor associating Sorafenib, method particularly includes: use selected by flow cytometry apoptosis NanogPos, then it is planted in 6 orifice plates, is treated that cell length is to 1 × 106Time, infecting slow virus Lv-sh-SIRT1 and slow virus Lv-sh-scramble, be expelled to NOD/SCID mouse after 72 hours respectively subcutaneous, cell quantity is 1 × 106/ only, 20 points of each injection.Treat that hypodermic tumour length is to 50mm3Time, experiment NOD/SCID mouse is divided into 4 groups: (1) blank group, the liver-cancer stem cell experimental mouse of inoculation slow virus Lv-sh-scramble, does not processes；(2) negative control group 1, the liver-cancer stem cell group experiment mice of inoculation slow virus Lv-sh-scramble, by 100mg/Kg gavage Sorafenib；(3) negative control group 2, the liver-cancer stem cell experiment mice of inoculation slow virus Lv-sh-SIRT1, do not process；(4) experimental group, the liver-cancer stem cell experiment mice of inoculation slow virus Lv-sh-SIRT1, by 100mg/Kg gavage Sorafenib；In experimentation, gavage group is every 3d gavage Sorafenib, and continuous processing puts to death mouse after 21 days, strips out tumour, measures gross tumor volume and tumor weight, and result is as shown in Figure 4.As shown in Figure 4, gavage Sorafenib can reduce the weight and volume of transplantable tumor, only interference SIRT1 expresses non-gavage Sorafenib and also can reduce the weight and volume of transplantable tumor, but the weight and volume effect that interference SIRT1 expresses associating Sorafenib process reduction transplantable tumor simultaneously is best, significant difference compared with blank group and control group.Result shows, interference SIRT1 expresses associating Sorafenib simultaneously and processes the growth that can suppress tumour, it is probably after interference SIRT1 expresses and reduces liver-cancer stem cell to Sorafenib drug resistance, utilize Sorafenib to kill liver-cancer stem cell, eventually reduce the weight and volume of transplantable tumor.
Transplantable tumor SABC: fix stripping out the paraformaldehyde that tumour mass fraction is 4%, FFPE, 4～5 μm serial section, examines under a microscope transplantable tumor pathology form after H&E dyes, then uses EnvisionTMImmunohistochemistry technique detects the expression of γ-H2AX, and Microscopic observation is also taken pictures, and result is as shown in Figure 5.As shown in Figure 5, H&E coloration result shows, compared with control group 1, interference SIRT1 expresses the transplantable tumor of associating Sorafenib process group, and its tumour cell form does not changes；Detection γ-H2AX finds that interference SIRT1 expresses the transplantable tumor damage compared with control group 1 and control group 2 of the transplantable tumor of associating Sorafenib process group and becomes apparent from.The studies above result further demonstrate that the relevant to Sorafenib resistance mechanism of SIRT1 and liver-cancer stem cell.
Four, the effect expressing connection Sorafenib of the SIRT1 Research Significance to clinical liver cancer treatment
The meaning of clinical liver cancer patient treatment is instructed for research SIRT1 associating Sorafenib further, the patient data that liver and gall surgical department of statistics Third Military Medical University southwest hospital liver cancer patient is treated from 2008 to 2010 years postoperative Nexavars, and follow up a case by regular visits to, find out simultaneously respective patient clinical pathology tissue specimen cut into slices, and with immunohistochemical staining detection SIRT1 expression.Statistics shows, has 53 example liver cancer patients not only with Sorafenib treatment but also there is clinical pathology tissue samples, but wherein only has 31 examples to have Follow-up Data.Then using immunohistochemical method to detect the expression of SIRT1 in this 31 example liver cancer patient paraffin specimen, detection method is as hereinbefore.Result shows: the positive rate that SIRT1 expresses in tumour is 64.5% (20/31), and negative rate is 36.5(11/31)；Using the relation of the expression life span postoperative with liver cancer patient of Kaplan-Meier methods analyst SIRT1, result is as shown in A in Fig. 6.From A in Fig. 6, expressing positive life span at liver cancer patient postoperative cancerous tissue SIRT1 and be substantially less than the patient that SIRT1 expression is negative, this is consistent in the expression statistics of 148 example liver cancer patient organization chips detection SIRT1 with before us.Using expressing of Kaplan-Meier methods analyst 31 example liver cancer patient SIRT1 to carry out treatment and the relation of prognosis with postoperative Sorafenib further, result is as shown in B in Fig. 6.Result shows, in cancerous tissue, SIRT1 expresses the positive, its life span of patient with Sorafenib treatment is short, and SIRT1 expression feminine gender Sorafenib treats patient in cancerous tissue, and its life span is significantly higher than cancerous tissue SIRT1 and expresses the patient of positive Sorafenib treatment.The above results indicates SIRT1 and expresses positive liver cancer patient, poor prognosis, treats insensitive with Sorafenib；It is obvious that SIRT1 expresses negative liver cancer patient Sorafenib result for the treatment of, and patient's good prognosis, life cycle is long.In sum, SIRT1 can be as an index of prediction patient's prognosis in liver cancer, and Sorafenib can be combined and instruct clinician's treatment to liver cancer patient, thus provide stronger theoretical foundation for treatment liver cancer and provide certain evidence for the drug target that research and development are new.
Finally illustrate is, preferred embodiment above is only in order to illustrate technical scheme and unrestricted, although the present invention being described in detail by above preferred embodiment, but skilled artisan would appreciate that, in the form and details it can be made various change, without departing from claims of the present invention limited range.
Claims (5)
- The application in the medicine of preparation suppression liver-cancer stem cell self of the 1.SIRT1 inhibitor associating Sorafenib.
- Application the most according to claim 1, it is characterised in that: described liver-cancer stem cell self is the propagation of liver-cancer stem cell, differentiation and Clone formation.
- 3. according to the application described in any one of claim 1-2, it is characterised in that: described SIRT1 inhibitor is Tenovin-6 or the slow virus of interference SIRT1 expression.
- Application the most according to claim 3, it is characterised in that: the slow virus that described interference SIRT1 expresses contains Lv-sh-SIRT1 plasmid.
- Application the most according to claim 4, it is characterised in that: the slow virus that described interference SIRT1 expresses is obtained through packaging, purifying by after Lv-sh-SIRT1 plasmid transfection 293T cell.
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| 去乙酰化酶SIRT1在肝癌干细胞自我更新中的作用及其机制研究;刘丽梅;《中国博士学位论文全文数据库医药卫生科技辑》;20140515(第5期);第E072-82页 * |
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