CN101869574B - 哇巴因在增强非小细胞肺癌细胞敏感性中的应用 - Google Patents
哇巴因在增强非小细胞肺癌细胞敏感性中的应用 Download PDFInfo
- Publication number
- CN101869574B CN101869574B CN2010102355289A CN201010235528A CN101869574B CN 101869574 B CN101869574 B CN 101869574B CN 2010102355289 A CN2010102355289 A CN 2010102355289A CN 201010235528 A CN201010235528 A CN 201010235528A CN 101869574 B CN101869574 B CN 101869574B
- Authority
- CN
- China
- Prior art keywords
- cell
- trail
- ouabain
- lung cancer
- tumor
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
- 208000002154 non-small cell lung carcinoma Diseases 0.000 title claims abstract description 41
- 208000029729 tumor suppressor gene on chromosome 11 Diseases 0.000 title claims abstract description 41
- 230000035945 sensitivity Effects 0.000 title description 14
- 230000001413 cellular effect Effects 0.000 title description 2
- 230000002708 enhancing effect Effects 0.000 title 1
- LPMXVESGRSUGHW-HBYQJFLCSA-N ouabain Chemical compound O[C@@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@H]1O[C@@H]1C[C@@]2(O)CC[C@H]3[C@@]4(O)CC[C@H](C=5COC(=O)C=5)[C@@]4(C)C[C@@H](O)[C@@H]3[C@@]2(CO)[C@H](O)C1 LPMXVESGRSUGHW-HBYQJFLCSA-N 0.000 claims description 76
- 244000166550 Strophanthus gratus Species 0.000 claims description 71
- LPMXVESGRSUGHW-UHFFFAOYSA-N Acolongiflorosid K Natural products OC1C(O)C(O)C(C)OC1OC1CC2(O)CCC3C4(O)CCC(C=5COC(=O)C=5)C4(C)CC(O)C3C2(CO)C(O)C1 LPMXVESGRSUGHW-UHFFFAOYSA-N 0.000 claims description 70
- LPMXVESGRSUGHW-GHYGWZAOSA-N Ouabain Natural products O([C@@H]1[C@@H](O)[C@@H](O)[C@@H](O)[C@H](C)O1)[C@H]1C[C@@H](O)[C@@]2(CO)[C@@](O)(C1)CC[C@H]1[C@]3(O)[C@@](C)([C@H](C4=CC(=O)OC4)CC3)C[C@@H](O)[C@H]21 LPMXVESGRSUGHW-GHYGWZAOSA-N 0.000 claims description 70
- 229960003343 ouabain Drugs 0.000 claims description 70
- 230000006907 apoptotic process Effects 0.000 claims description 21
- 230000008878 coupling Effects 0.000 claims description 11
- 238000010168 coupling process Methods 0.000 claims description 11
- 238000005859 coupling reaction Methods 0.000 claims description 11
- 230000006698 induction Effects 0.000 claims description 5
- 238000002360 preparation method Methods 0.000 claims description 4
- 108060008682 Tumor Necrosis Factor Proteins 0.000 claims description 3
- 239000003446 ligand Substances 0.000 claims description 3
- 102000003390 tumor necrosis factor Human genes 0.000 claims description 3
- 239000003560 cancer drug Substances 0.000 claims 1
- 239000002532 enzyme inhibitor Substances 0.000 abstract description 6
- 229940125532 enzyme inhibitor Drugs 0.000 abstract description 6
- 239000002246 antineoplastic agent Substances 0.000 abstract description 3
- 229940041181 antineoplastic drug Drugs 0.000 abstract description 3
- BITYAPCSNKJESK-UHFFFAOYSA-N potassiosodium Chemical compound [Na].[K] BITYAPCSNKJESK-UHFFFAOYSA-N 0.000 abstract description 2
- 210000004027 cell Anatomy 0.000 description 85
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 21
- 239000003814 drug Substances 0.000 description 18
- 230000000694 effects Effects 0.000 description 16
- 210000004881 tumor cell Anatomy 0.000 description 15
- 206010028980 Neoplasm Diseases 0.000 description 12
- 229940079593 drug Drugs 0.000 description 12
- 238000005406 washing Methods 0.000 description 11
- 101000610604 Homo sapiens Tumor necrosis factor receptor superfamily member 10B Proteins 0.000 description 10
- 230000000259 anti-tumor effect Effects 0.000 description 10
- 102000004190 Enzymes Human genes 0.000 description 8
- 108090000790 Enzymes Proteins 0.000 description 8
- 229940088598 enzyme Drugs 0.000 description 8
- 102000003952 Caspase 3 Human genes 0.000 description 7
- 108090000397 Caspase 3 Proteins 0.000 description 7
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 7
- 201000011510 cancer Diseases 0.000 description 7
- 201000005202 lung cancer Diseases 0.000 description 7
- 208000020816 lung neoplasm Diseases 0.000 description 7
- 229940097217 cardiac glycoside Drugs 0.000 description 6
- 239000002368 cardiac glycoside Substances 0.000 description 6
- 210000000170 cell membrane Anatomy 0.000 description 6
- 239000012531 culture fluid Substances 0.000 description 6
- 230000004927 fusion Effects 0.000 description 6
- 229930002534 steroid glycoside Natural products 0.000 description 6
- 241000699660 Mus musculus Species 0.000 description 5
- 230000001640 apoptogenic effect Effects 0.000 description 5
- 230000034994 death Effects 0.000 description 5
- MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical compound O1C(=O)C2=CC(N=C=S)=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 description 5
- 230000014509 gene expression Effects 0.000 description 5
- 238000011580 nude mouse model Methods 0.000 description 5
- 102000004121 Annexin A5 Human genes 0.000 description 4
- 108090000672 Annexin A5 Proteins 0.000 description 4
- 240000001879 Digitalis lutea Species 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- 230000001939 inductive effect Effects 0.000 description 4
- 238000002372 labelling Methods 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- 102000004169 proteins and genes Human genes 0.000 description 4
- 108090000623 proteins and genes Proteins 0.000 description 4
- 230000004614 tumor growth Effects 0.000 description 4
- FWBHETKCLVMNFS-UHFFFAOYSA-N 4',6-Diamino-2-phenylindol Chemical compound C1=CC(C(=N)N)=CC=C1C1=CC2=CC=C(C(N)=N)C=C2N1 FWBHETKCLVMNFS-UHFFFAOYSA-N 0.000 description 3
- LTMHDMANZUZIPE-AMTYYWEZSA-N Digoxin Natural products O([C@H]1[C@H](C)O[C@H](O[C@@H]2C[C@@H]3[C@@](C)([C@@H]4[C@H]([C@]5(O)[C@](C)([C@H](O)C4)[C@H](C4=CC(=O)OC4)CC5)CC3)CC2)C[C@@H]1O)[C@H]1O[C@H](C)[C@@H](O[C@H]2O[C@@H](C)[C@H](O)[C@@H](O)C2)[C@@H](O)C1 LTMHDMANZUZIPE-AMTYYWEZSA-N 0.000 description 3
- 206010027336 Menstruation delayed Diseases 0.000 description 3
- 208000005718 Stomach Neoplasms Diseases 0.000 description 3
- 238000002512 chemotherapy Methods 0.000 description 3
- LTMHDMANZUZIPE-PUGKRICDSA-N digoxin Chemical compound C1[C@H](O)[C@H](O)[C@@H](C)O[C@H]1O[C@@H]1[C@@H](C)O[C@@H](O[C@@H]2[C@H](O[C@@H](O[C@@H]3C[C@@H]4[C@]([C@@H]5[C@H]([C@]6(CC[C@@H]([C@@]6(C)[C@H](O)C5)C=5COC(=O)C=5)O)CC4)(C)CC3)C[C@@H]2O)C)C[C@@H]1O LTMHDMANZUZIPE-PUGKRICDSA-N 0.000 description 3
- 229960005156 digoxin Drugs 0.000 description 3
- LTMHDMANZUZIPE-UHFFFAOYSA-N digoxine Natural products C1C(O)C(O)C(C)OC1OC1C(C)OC(OC2C(OC(OC3CC4C(C5C(C6(CCC(C6(C)C(O)C5)C=5COC(=O)C=5)O)CC4)(C)CC3)CC2O)C)CC1O LTMHDMANZUZIPE-UHFFFAOYSA-N 0.000 description 3
- 238000009826 distribution Methods 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 206010017758 gastric cancer Diseases 0.000 description 3
- 230000012010 growth Effects 0.000 description 3
- 230000012447 hatching Effects 0.000 description 3
- 238000011081 inoculation Methods 0.000 description 3
- JLPDBLFIVFSOCC-XYXFTTADSA-N oleandrin Chemical compound O1[C@@H](C)[C@H](O)[C@@H](OC)C[C@@H]1O[C@@H]1C[C@@H](CC[C@H]2[C@]3(C[C@@H]([C@@H]([C@@]3(C)CC[C@H]32)C=2COC(=O)C=2)OC(C)=O)O)[C@]3(C)CC1 JLPDBLFIVFSOCC-XYXFTTADSA-N 0.000 description 3
- 229950010050 oleandrin Drugs 0.000 description 3
- 238000001556 precipitation Methods 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 201000011549 stomach cancer Diseases 0.000 description 3
- 230000001225 therapeutic effect Effects 0.000 description 3
- 231100000331 toxic Toxicity 0.000 description 3
- 230000002588 toxic effect Effects 0.000 description 3
- 231100000419 toxicity Toxicity 0.000 description 3
- 230000001988 toxicity Effects 0.000 description 3
- 238000012546 transfer Methods 0.000 description 3
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 2
- 208000017897 Carcinoma of esophagus Diseases 0.000 description 2
- 206010057248 Cell death Diseases 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- WDJUZGPOPHTGOT-OAXVISGBSA-N Digitoxin Natural products O([C@H]1[C@@H](C)O[C@@H](O[C@@H]2C[C@@H]3[C@@](C)([C@@H]4[C@H]([C@]5(O)[C@@](C)([C@H](C6=CC(=O)OC6)CC5)CC4)CC3)CC2)C[C@H]1O)[C@H]1O[C@@H](C)[C@H](O[C@H]2O[C@@H](C)[C@@H](O)[C@@H](O)C2)[C@@H](O)C1 WDJUZGPOPHTGOT-OAXVISGBSA-N 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- -1 FLIIP Proteins 0.000 description 2
- 102100034343 Integrase Human genes 0.000 description 2
- 206010022998 Irritability Diseases 0.000 description 2
- 241000699670 Mus sp. Species 0.000 description 2
- 206010030155 Oesophageal carcinoma Diseases 0.000 description 2
- JLPDBLFIVFSOCC-UHFFFAOYSA-N Oleandrin Natural products O1C(C)C(O)C(OC)CC1OC1CC(CCC2C3(CC(C(C3(C)CCC32)C=2COC(=O)C=2)OC(C)=O)O)C3(C)CC1 JLPDBLFIVFSOCC-UHFFFAOYSA-N 0.000 description 2
- 108010092799 RNA-directed DNA polymerase Proteins 0.000 description 2
- FKNQFGJONOIPTF-UHFFFAOYSA-N Sodium cation Chemical compound [Na+] FKNQFGJONOIPTF-UHFFFAOYSA-N 0.000 description 2
- 208000027418 Wounds and injury Diseases 0.000 description 2
- 230000004913 activation Effects 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 239000002299 complementary DNA Substances 0.000 description 2
- 239000000470 constituent Substances 0.000 description 2
- 231100000135 cytotoxicity Toxicity 0.000 description 2
- 230000003013 cytotoxicity Effects 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- WDJUZGPOPHTGOT-XUDUSOBPSA-N digitoxin Chemical compound C1[C@H](O)[C@H](O)[C@@H](C)O[C@H]1O[C@@H]1[C@@H](C)O[C@@H](O[C@@H]2[C@H](O[C@@H](O[C@@H]3C[C@@H]4[C@]([C@@H]5[C@H]([C@]6(CC[C@@H]([C@@]6(C)CC5)C=5COC(=O)C=5)O)CC4)(C)CC3)C[C@@H]2O)C)C[C@@H]1O WDJUZGPOPHTGOT-XUDUSOBPSA-N 0.000 description 2
- 229960000648 digitoxin Drugs 0.000 description 2
- 231100000673 dose–response relationship Toxicity 0.000 description 2
- 201000005619 esophageal carcinoma Diseases 0.000 description 2
- 239000000284 extract Substances 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 230000003301 hydrolyzing effect Effects 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- 238000011835 investigation Methods 0.000 description 2
- 230000001665 lethal effect Effects 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 231100000252 nontoxic Toxicity 0.000 description 2
- 230000003000 nontoxic effect Effects 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 108020003175 receptors Proteins 0.000 description 2
- 102000005962 receptors Human genes 0.000 description 2
- 238000010839 reverse transcription Methods 0.000 description 2
- 229910001415 sodium ion Inorganic materials 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 238000005728 strengthening Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 208000024891 symptom Diseases 0.000 description 2
- 231100000820 toxicity test Toxicity 0.000 description 2
- AOUOVFRSCMDPFA-QSDJMHMYSA-N (2s)-2-[[(2s)-2-[[(2s)-2-[[(2s)-2-amino-3-carboxypropanoyl]amino]-4-carboxybutanoyl]amino]-3-methylbutanoyl]amino]butanedioic acid Chemical compound OC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](N)CC(O)=O AOUOVFRSCMDPFA-QSDJMHMYSA-N 0.000 description 1
- 208000010507 Adenocarcinoma of Lung Diseases 0.000 description 1
- 241000208327 Apocynaceae Species 0.000 description 1
- 108010063104 Apoptosis Regulatory Proteins Proteins 0.000 description 1
- 102000010565 Apoptosis Regulatory Proteins Human genes 0.000 description 1
- 102100021569 Apoptosis regulator Bcl-2 Human genes 0.000 description 1
- 241000219193 Brassicaceae Species 0.000 description 1
- 206010007559 Cardiac failure congestive Diseases 0.000 description 1
- 241000208365 Celastraceae Species 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
- 108090000695 Cytokines Proteins 0.000 description 1
- 108020004414 DNA Proteins 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 108090000288 Glycoproteins Proteins 0.000 description 1
- 102000003886 Glycoproteins Human genes 0.000 description 1
- 206010019280 Heart failures Diseases 0.000 description 1
- 101000971171 Homo sapiens Apoptosis regulator Bcl-2 Proteins 0.000 description 1
- 101000610602 Homo sapiens Tumor necrosis factor receptor superfamily member 10C Proteins 0.000 description 1
- 101000610609 Homo sapiens Tumor necrosis factor receptor superfamily member 10D Proteins 0.000 description 1
- 206010025323 Lymphomas Diseases 0.000 description 1
- 206010027476 Metastases Diseases 0.000 description 1
- ZDZOTLJHXYCWBA-VCVYQWHSSA-N N-debenzoyl-N-(tert-butoxycarbonyl)-10-deacetyltaxol Chemical compound O([C@H]1[C@H]2[C@@](C([C@H](O)C3=C(C)[C@@H](OC(=O)[C@H](O)[C@@H](NC(=O)OC(C)(C)C)C=4C=CC=CC=4)C[C@]1(O)C3(C)C)=O)(C)[C@@H](O)C[C@H]1OC[C@]12OC(=O)C)C(=O)C1=CC=CC=C1 ZDZOTLJHXYCWBA-VCVYQWHSSA-N 0.000 description 1
- 206010028851 Necrosis Diseases 0.000 description 1
- 108091093105 Nuclear DNA Proteins 0.000 description 1
- 206010030113 Oedema Diseases 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 229930012538 Paclitaxel Natural products 0.000 description 1
- 108010019160 Pancreatin Proteins 0.000 description 1
- 241000013557 Plantaginaceae Species 0.000 description 1
- NPYPAHLBTDXSSS-UHFFFAOYSA-N Potassium ion Chemical compound [K+] NPYPAHLBTDXSSS-UHFFFAOYSA-N 0.000 description 1
- 206010060862 Prostate cancer Diseases 0.000 description 1
- 102000006382 Ribonucleases Human genes 0.000 description 1
- 108010083644 Ribonucleases Proteins 0.000 description 1
- 102000000763 Survivin Human genes 0.000 description 1
- 108010002687 Survivin Proteins 0.000 description 1
- 108010000449 TNF-Related Apoptosis-Inducing Ligand Receptors Proteins 0.000 description 1
- 102000002259 TNF-Related Apoptosis-Inducing Ligand Receptors Human genes 0.000 description 1
- 108091008005 TRAIL–DR complexes Proteins 0.000 description 1
- 102100040115 Tumor necrosis factor receptor superfamily member 10C Human genes 0.000 description 1
- 102100040110 Tumor necrosis factor receptor superfamily member 10D Human genes 0.000 description 1
- 206010052428 Wound Diseases 0.000 description 1
- 210000000683 abdominal cavity Anatomy 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000036982 action potential Effects 0.000 description 1
- 239000011543 agarose gel Substances 0.000 description 1
- 238000000246 agarose gel electrophoresis Methods 0.000 description 1
- 238000003915 air pollution Methods 0.000 description 1
- 238000000137 annealing Methods 0.000 description 1
- 230000000118 anti-neoplastic effect Effects 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 239000012930 cell culture fluid Substances 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 229960004316 cisplatin Drugs 0.000 description 1
- DQLATGHUWYMOKM-UHFFFAOYSA-L cisplatin Chemical compound N[Pt](N)(Cl)Cl DQLATGHUWYMOKM-UHFFFAOYSA-L 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 229940127089 cytotoxic agent Drugs 0.000 description 1
- 239000002254 cytotoxic agent Substances 0.000 description 1
- 238000007405 data analysis Methods 0.000 description 1
- 230000007850 degeneration Effects 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 238000006477 desulfuration reaction Methods 0.000 description 1
- 230000023556 desulfurization Effects 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000007599 discharging Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000002934 diuretic Substances 0.000 description 1
- 230000001882 diuretic effect Effects 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- 239000003792 electrolyte Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 108010048367 enhanced green fluorescent protein Proteins 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 210000003527 eukaryotic cell Anatomy 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000000684 flow cytometry Methods 0.000 description 1
- 238000000799 fluorescence microscopy Methods 0.000 description 1
- 238000001943 fluorescence-activated cell sorting Methods 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 208000019622 heart disease Diseases 0.000 description 1
- 238000010166 immunofluorescence Methods 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 239000002440 industrial waste Substances 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 230000037427 ion transport Effects 0.000 description 1
- 230000002147 killing effect Effects 0.000 description 1
- 230000001795 light effect Effects 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 206010025482 malaise Diseases 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000010534 mechanism of action Effects 0.000 description 1
- 238000002483 medication Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000002107 myocardial effect Effects 0.000 description 1
- 230000017074 necrotic cell death Effects 0.000 description 1
- 210000004498 neuroglial cell Anatomy 0.000 description 1
- 210000002569 neuron Anatomy 0.000 description 1
- 230000002018 overexpression Effects 0.000 description 1
- 229960001592 paclitaxel Drugs 0.000 description 1
- 229940055695 pancreatin Drugs 0.000 description 1
- 239000008194 pharmaceutical composition Substances 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 229910001414 potassium ion Inorganic materials 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 230000000284 resting effect Effects 0.000 description 1
- 238000003757 reverse transcription PCR Methods 0.000 description 1
- 238000012552 review Methods 0.000 description 1
- 239000003161 ribonuclease inhibitor Substances 0.000 description 1
- 229920002477 rna polymer Polymers 0.000 description 1
- 230000000391 smoking effect Effects 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- RCINICONZNJXQF-MZXODVADSA-N taxol Chemical compound O([C@@H]1[C@@]2(C[C@@H](C(C)=C(C2(C)C)[C@H](C([C@]2(C)[C@@H](O)C[C@H]3OC[C@]3([C@H]21)OC(C)=O)=O)OC(=O)C)OC(=O)[C@H](O)[C@@H](NC(=O)C=1C=CC=CC=1)C=1C=CC=CC=1)O)C(=O)C1=CC=CC=C1 RCINICONZNJXQF-MZXODVADSA-N 0.000 description 1
- 229940063683 taxotere Drugs 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 230000026683 transduction Effects 0.000 description 1
- 238000010361 transduction Methods 0.000 description 1
- 230000032895 transmembrane transport Effects 0.000 description 1
- 210000005239 tubule Anatomy 0.000 description 1
- 230000007306 turnover Effects 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Images
Landscapes
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
本发明涉及钠钾ATP酶抑制剂哇巴因在逆转非小细胞肺癌细胞对靶向性抗肿瘤药物耐受的新用途。具体是哇巴因可作为治疗非小细胞肺癌的增敏剂,与TRAIL联合使用用于非小细胞肺癌的治疗。
Description
技术领域
本发明涉及钠钾ATP酶抑制剂哇巴因在逆转非小细胞肺癌细胞对靶向性抗肿瘤药物耐受的新用途。
背景技术
恶性肿瘤是危害人类健康的重大疾病。江苏省近几年开展了以恶性肿瘤为主的全人口死因回顾性调查,结果显示男女性第1位死因均为恶性肿瘤,其死亡率占全部死因的27.41%。资料显示,2003年江苏省城市地区位于恶性肿瘤死亡前4位的分别是肺癌、胃癌、肝癌、食管癌;而农村地区为肝癌、肺癌、食管癌、胃癌。肺癌和胃癌死亡率城市明显高于农村。尤其是通过分析1973至2004年几种主要恶性肿瘤的死亡情况发现,肺癌死亡率大幅上升,2004年肺癌死亡率是1973年的5.65倍。由此可见,肺癌已经成为我省恶性肿瘤数一数二的“杀手”,严重威胁着我省人民的生存健康。
为什么在短短的几十年间,肺癌的发病率及死亡率有如此迅猛的发展趋势?1:大样品调查研究表明肺癌的主要危险因素为吸烟和大气污染,而近年来吸烟人数有增加的趋势,汽车尾气和工业废气的大量排放,又加重了大气环境污染的程度。因此,随着我省经济的发展,城市化、工业化进程的加快,空气污染将会越来越严重,这将不可避免地导致肺癌死亡率进一步增加。2:肺癌在早期时往往没有明显的症状,当有症状被诊断出来时,常常已经是局部晚期或晚期无法手术切除的情形。一般而言,新诊断的肺癌仅约不到20%的病人属于早期可以有机会接受手术切除的病患。而另外80%的晚期肺癌病人则失去了外科手术根治的最佳时机。3:到目前为止,对肺癌的治疗,尤其是晚期肺癌病人缺乏非常有效的治疗干预手段。
临床上,肺癌通常分为小细胞与非小细胞肺癌两种类型,总的肺癌病例中,非小细胞肺癌(包括肺鳞癌,肺腺癌)约占80%。因此针对非小细胞肺癌的治疗研究是攻克肺癌的一项极其重要的课题,引起了全世界医学界的广泛关注。
如上所述,虽然手术治疗是治疗非小细胞肺癌的重要手段,但对绝大部分晚期肺癌病人来说手术治疗并不能有效解决肿瘤转移的问题。因此对于这些晚期或已经有远端转移的肺癌病人,全身性的化学治疗是非小细胞肺癌多学科综合治疗的主要策略之一。尤其是化疗与手术、放射等疗法综合运用能明显防止癌肿转移、复发,提高长期生存率。现在,非小细胞肺癌化疗已经初步形成共识,通常采用顺铂加泰索帝、紫杉醇、健择、诺维本等中的一种。根据患者的具体情况,选择不同的组合,可以达到提高疗效,降低毒性反应的目的,特别是出现了一些极具前景的新型靶向药物,其作用机制与传统化疗药物不同。传统化疗药物属于细胞毒性药物,是通过毒性杀死肿瘤细胞,但同时不可避免会伤害正常细胞。而靶向药物进入肿瘤细胞后,可以通过特异性作用于肿瘤生长的细胞信号途径,抑制肿瘤细胞的增殖、浸润、转移,不良反应轻,患者可以很好耐受。
在这些靶向药物中,TRAIL(肿瘤坏死因子相关的凋亡诱导配体)越来越受到人们的关注。TRAIL是由Wiley于1995年检索EST时首次发现并克隆的,是一种II型糖蛋白,属于肿瘤坏死因子家族,与相应受体结合后,可以快速诱导细胞发生凋亡。但与其它凋亡诱导分子显著不同的是,TRAIL主要杀伤转化细胞和肿瘤细胞,而正常细胞可以逃逸它的杀伤作用。体内研究表明,TRAIL可以明显抑制肿瘤生长,甚至彻底清除肿瘤。TRAIL的这种肿瘤细胞选择性杀伤作用使该蛋白在抗肿瘤方面具有很强的开发价值与潜力。但是值得注意的是,并非所有的肿瘤细胞对TRAIL都具有很强的敏感性。体外研究表明,有些肿瘤细胞株对TRAIL诱导凋亡作用具有很强的耐受性。这可能与肿瘤细胞表面缺乏TRAIL受体(DR4,DR5),或分布有假受体(DcR1,DcR2),或过度表达一些抗凋亡蛋白如bcl-2,FLIIP,survivin有关。因此TRAIL的抗肿瘤作用因不同的肿瘤类型而异。
研究表明,非小细胞肺癌细胞,如A549对TRAIL有明显的耐受性,但具体的机理目前并不清楚。为了充分发挥TRAIL对肿瘤细胞的选择杀伤作用,有必要寻找新的策略以逆转非小细胞肺癌细胞株对TRAIL的敏感性,从而达到运用TRAIL有效治疗非小细胞肺癌的目的。而作为一个理想干预手段必须具备以下特点:1:显著性增强TRAIL对非小细胞肺癌细胞的细胞凋亡诱导作用;2:对非小细胞肺癌细胞具有高度的选择性;即不会对正常细胞及机体产生较强的非特异性的毒副作用。事实上,虽然有些化疗药物能明显逆转非小细胞肺癌细胞株A549对TRAIL的敏感性,但这些化疗药物本身对肿瘤细胞的选择性不够强,因此常常以造成对正常组织与细胞的损伤为代价。
钠钾ATP酶是分布于真核生物细胞膜重要的离子转运泵,通过消耗ATP能量将钠离子与钾离子逆浓度梯度进行跨膜转运,从而维持细胞膜内外稳定的钠钾离子浓度梯度、细胞膜静息电位与细胞渗透压平衡。在兴奋性细胞如神经元细胞或心肌细胞中,钠钾ATP酶与动作电位产生、兴奋信号传递密切相关。在非兴奋性细胞如肾小管上皮细胞中,钠钾ATP酶是肾小管对水分、钠离子进行重吸收、营养物质摄取的关键蛋白,对维持机体电解质平衡、酸碱平衡、营养物质正常利用发挥至关重要的作用,鉴于此,机体平均1/3ATP能量用于维持该泵的正常运转。
从天然植物洋地黄中提取的强心苷成分是钠钾ATP酶抑制剂与细胞膜钠钾ATP酶α亚基有很强的亲合力,结合后通过构象改变抑制钠钾ATP酶对Na+,K+离子的转运能力,具有多种生物活性。自1785年W.Withering使用紫花洋地黄叶(Digitalis purpurea)治疗水肿以来,至今已经在夹竹桃科、玄参科、十字花科、卫矛科等10几个科的100多种植物中发现强心苷,常见的较重要的有洋地黄毒苷(Digitoxin),异羟基洋地黄毒苷(Digoxin),夹竹桃苷(Oleandrin),乌本苷(ouabain)等。中药洋地黄类成分长期以来一直用于治疗充血性心力衰竭及节律性障碍等心脏疾病,但同时该类成分也具有明显的抗肿瘤效果。研究表明,强心苷如digoxin,oleandrin以及ouabain能诱导多种肿瘤细胞,如淋巴瘤细胞、前列腺癌细胞、神经胶质瘤细胞的凋亡或坏死,从而具有明显的抗肿瘤效果。
强心苷的这种抗肿瘤作用通常认为是由强心苷的细胞毒性引起的,因此人们对钠钾ATP酶物质是否能用于肿瘤治疗产生怀疑。但是越来越多的研究表明,强心苷,如哇巴因在非毒性剂量下也能显著性抑制肿瘤细胞的生长和转移。遵循这样的思路,我们在前期的工作中发现钠钾ATP酶抑制剂哇巴因能在非毒性剂量(20-150nM)下很显著性地增强非小细胞肺癌细胞对TRAIL的敏感性,从而逆转非小细胞肺癌细胞对TRAIL的耐受性。因此本研究发明钠钾ATP酶抑制剂在抗肿瘤药物制备中的新用途。
发明内容
本发明的目的是提供钠钾ATP酶抑制剂在抗肿瘤药物制备中的新用途,即作为抗肿瘤增敏剂,该增敏剂能能显著逆转非小细胞肺癌细胞对TRAIL耐受性。
本发明公开了哇巴因在增强非小细胞肺癌细胞对肿瘤坏死因子相关的凋亡诱导配体(TRAIL)敏感性中的应用。
基于上述应用,哇巴因可作为治疗非小细胞肺癌的增敏剂,与TRAIL联合使用用于非小细胞肺癌的治疗。
哇巴因也可以与TRAIL一起制成药物组合物,从而在制备治疗非小细胞肺癌的药物中应用。
上述所说的应用中,哇巴因的剂量为(20-150nM)。
本发明中经实验显示,哇巴因能显著性提高非小细胞肺癌细胞膜DR4与DR5mRNA的表达量,上调DR4与DR5可能是哇巴因增强TRAIL诱导的A549细胞凋亡的重要手段。在毒性实验中显示,在明显诱导A549细胞凋亡的情况下,TRAIL+哇巴因对正常细胞无明显的毒性。
通过细胞水平与裸鼠实验,表明TRAIL与哇巴因的药物组合能使非小细胞肺癌细胞凋亡率接近90%,体内抗肿瘤实验结果表明这两种药的组合使肺癌抑制率达到70%以上。因此哇巴因除了具有强心利尿的传统药理作用外,尚能作为效果很好的抗肿瘤增敏剂。
附图说明
图1:哇巴因增强非小细胞肺癌细胞A549对TRAIL的敏感性图2:哇巴因剂量依赖性地增强非小细胞肺癌细胞A549对TRAIL的敏感性图3:哇巴因联用TRAIL对非小细胞肺癌细胞A549核碎裂图4:哇巴因联用TRAIL对非小细胞肺癌细胞A549caspase3活性激活图5:哇巴因选择性增强非小细胞肺癌细胞对TRAIL的敏感性图6:哇巴因上调A549细胞表面死亡受体DR4与DR5mRNA表达水平图7:哇巴因上调A549细胞表面死亡受体DR4与DR5分布图8:哇巴因与TRAIL联用抑制裸鼠接种A549肿瘤生长曲线图9:哇巴因与TRAIL联合应用抑制裸鼠接种A549肿瘤生长图。
具体实施方式
实验材料哇巴因、DAPI、PI、DEPC、Trizol购自Sigma公司,A549非小细胞肺癌细胞株购自ATCC公司。DMEM细胞培养液、胰酶购自美国Hyclone公司。caspase3检测试剂盒购自Biovision公司。EGFP-Annexin V为南京大学医药生物技术国家重点实验室表达。流式细胞仪FACS Calibur(美国Becton Dickson公司),DR4与DR5抗体购自美国Milipore公司,半干半湿法蛋白质转膜仪(美国Biorad公司),DNA凝胶电泳仪(南京大学电子设备厂)。逆转录体系dNTP、oligo(dT)18、RNase inhibitor、逆转录酶、PCR试剂盒均购自Takara公司。琼脂糖凝胶、EB购自南京大治生物公司。LipofectamineTM2000购自美国Invitrogen公司。
实施例一Annexin V/PI流式细胞仪法检测TRAIL与哇巴因联用致A549细胞凋亡非小细胞肺癌细胞A549于DMEM+10%胎牛血清培养液中培养,培养条件为37℃,5%CO2的湿润无菌环境。细胞长至80%-90%融合时,用PBS洗涤2次,0.25%胰酶消化。按细胞量6×104/孔接种于24孔细胞培养板中。待细胞贴壁生长完全后,加入药物分别为TRAIL10ng/ml,TRAIL 10ng/ml+哇巴因100nM,TRAIL 20ng/ml,TRAIL 20ng/ml+哇巴因100nM,TRAIL 50ng/ml,TRAIL 50ng/ml+哇巴因100nM,TRAIL 100ng/ml,TRAIL100ng/ml+哇巴因100nM,TRAIL 200ng/ml,TRAIL 200ng/ml+哇巴因100nM,TRAIL400ng/ml,TRAIL 400ng/ml+哇巴因100nM,每组3个孔(图1),或分别为TRAIL100ng/ml+哇巴因10nM,哇巴因10nM,TRAIL 100ng/ml+哇巴因20nM,哇巴因20nM,TRAIL100ng/ml+哇巴因30nM,哇巴因30nM,TRAIL 100ng/ml+哇巴因50nM,哇巴因50nM,TRAIL100ng/ml+哇巴因100nM,哇巴因100nM,每组3个孔(图2)。
细胞经药物处理12h后收集,用预冷PBS洗2遍,加入EGFP标记的Annexin V(2μL),冰上孵育20min,迅速加入PI(1μg/mL),于流式细胞仪上检测细胞凋亡情况,Annexin V(+)/PI(-)为早期凋亡细胞。
结果:采用Annexin V/PI双染法考察A549细胞对TRAIL的敏感性,发现只有在使用TRAIL达到400ng/ml时,才使得A549细胞有轻微的细胞凋亡发生,表明该细胞对TRAIL并不敏感(图1)。同样,哇巴因在使用浓度到100nM时,本身对肿瘤细胞具有轻微的细胞凋亡作用(图2),但是两药的联用能极显著性地增强A549细胞对细胞凋亡的敏感性,从图1中可知,在哇巴因使用浓度为100nM时,TRAIL能剂量依赖性地增强A549细胞凋亡,在TRAIL使用浓度达到100ng/ml时,已经能使细胞凋亡达到75%以上,表明TRAIL与哇巴因组合是很有前景的药物组合。同样地,从图2中可知,当TRAIL使用浓度为100ng/ml时,哇巴因也能剂量性地增强A549细胞凋亡。
实施例二荧光显微镜法检测TRAIL与哇巴因联用致A549细胞核碎裂非小细胞肺癌细胞A549于DMEM+10%胎牛血清培养液中培养,培养条件为37℃,5%CO2的湿润无菌环境。细胞长至80%-90%融合时,用PBS洗涤2次,0.25%胰酶消化。按细胞量1×105/孔接种于6孔细胞培养板中。培养板中预先放置灭菌过的盖玻片面,待细胞贴壁生长至60%时,加入药物处理,分别为对照组、TRAIL 100ng/ml,TRAIL 100ng/ml+哇巴因50nM,TRAIL 100ng/ml,TRAIL 100ng/ml+哇巴因50nM,每组重复3个孔(图3)。处理12h后,将载玻片取出,冷PBS洗涤2次,滴加DAPI(1μg/ml)数滴后于荧光显微镜上观察,拍片。
结果:DAPI是一种常见的细胞核染料,能掺入到核DNA中,在激发光作用下,呈现蓝色核,通过观察发现(图3),单用哇巴因50nM与TRAIL100ng/ml时,细胞核相对完整,没有明显的细胞凋亡核碎裂,但是当TRAIL与哇巴因联用时,细胞核破碎明显,进一步表明哇巴因能增强A549细胞对TRAIL的敏感性。
实施例三TRAIL与哇巴因对caspase3水解酶活性的影响非小细胞肺癌细胞A549于DMEM+10%胎牛血清培养液中培养,培养条件为37℃,5%CO2的湿润无菌环境。细胞长至80%-90%融合时,用PBS洗涤2次,0.25%胰酶消化。按细胞量6×104/孔接种于24孔细胞培养板中。加入药物处理,分别为对照组、TRAIL100ng/ml,TRAIL 100ng/ml+哇巴因50nM,TRAIL 100ng/ml,TRAIL 100ng/ml+哇巴因50nM,每组重复3个孔(图4)。处理12h后,将细胞用胰酶消化后,加入FITC标记的DEVD.fmk 1-2μl,于冰上孵育30min后,PBS洗2遍,流式细胞仪分析FITC荧光强度。
结果:当细胞处于凋亡状态时,会引起caspase3的激活,从而使细胞中诸多蛋白质发生非选择性切割,最终使细胞走向凋亡。采用流式细胞仪方法考察TRAIL与哇巴因对caspase3水解酶活性的影响,底物为FITC-DEVE.fmk。其基本原理在于当细胞内caspase3酶激活时,会切割FITC-DEVD.fmk,从而使细胞内荧光能在488nm激发光激发下被检测到(图4)。结果与免疫荧光相符,在单用TRAIL或哇巴因情况下,细胞内caspase3酶激活不明显,但是两药的联用,细胞内caspase3被激活。
实施例四毒性实验HEK293与A549细胞分别于DMEM+10%胎牛血清培养液中培养,培养条件为37℃,5%CO2的湿润无菌环境。细胞长至80%-90%融合时,用PBS洗涤2次,0.25%胰酶消化。按细胞量5×103/孔接种于96孔细胞培养板中。加入药物TRAIL 100ng/ml+哇巴因100nM处理0,8,12,24,48h,每组重复3个孔(图5),处理12h后,收集细胞,按实施例一检测A549与HEK293细胞凋亡。
结果:为考察TRAIL与哇巴因的联用增效作用是否对正常细胞具有毒性副作用。本研究比较TRAIL100ng/ml+哇巴因(100nM)在正常细胞株(HEK293)及肿瘤细胞株(A549)上的细胞毒性差异。结果见图5,在明显诱导A549细胞凋亡的情况下,TRAIL+哇巴因对正常细胞无明显的毒性,这些数据说明TRAIL+哇巴因对肿瘤具有明显的选择性,是具有开发前景的治疗方案。
实施例五哇巴因对A549细胞DR4与DR5mRNA与蛋白质表达的影响1、细胞mRNA的提取所有物品均提前用DEPC处理并灭菌。非小细胞肺癌细胞A549于DMEM+10%胎牛血清培养液中培养,培养条件为37℃,5%CO2的湿润无菌环境。细胞长至80%-90%融合时,用PBS洗涤2次,0.25%胰酶消化。按细胞量1×105/孔接种于6孔细胞培养板中。铺细胞于6孔板,等细胞长到80%-90%后,加药(分别为哇巴因0,50,100,200nM),处理12h后收集细胞,PBS洗涤一次。加入1mL TRIZOL,用枪吹打至细胞完全溶解,室温放置5min。加入200μL的氯仿,振荡15s后放置2-3min,2-8℃12000g离心15min。吸取上层,转移到干净eppendorf管中,加入500μL异丙醇,放置10min,2-8℃下12000g离心10min,可见RNA沉淀。弃上清,加入1mL75%乙醇洗涤沉淀,7500g离心5min,在空气中晾干RNA沉淀,加入30μL DEPC处理的DDW溶解。
2.、逆转录PCR采用20μL逆转录体系,将提取的模板RNA约1μg与5×buffer4μL,dNTP(10mM)2μL,oligo(dT)18(10μmol/L)1μL,RNase Inhibitor1μL,逆转录酶1μL混匀后30℃水浴10min,42℃水浴30min,98℃水浴10min。取等量的cDNA做模板做PCR扩增,采用20μL PCR反应体系:DDW15.2μL,10×Taq buffer2.5μL,dNTP2μL,Mg2+2μL,上游引物1μL,下游引物1μL,cDNA模板1μL,Taq polymerase0.3μL。PCR条件:94℃热变性4min后;94℃变性40s,55℃退火40s,72℃延伸40s(循环数为20-45个);最后72℃延伸7min。PCR产物进行1%琼脂糖凝胶电泳,EB显色。
3、流式细胞仪检测细胞膜DR4与DR5分布非小细胞肺癌细胞A549于DMEM+10%胎牛血清培养液中培养,培养条件为37℃,5%CO2的湿润无菌环境。细胞长至80%-90%融合时,用PBS洗涤2次,0.25%胰酶消化。按细胞量1×105/孔接种于6孔细胞培养板中。收集药物处理后细胞,PBS洗涤2遍后,加入含2%BSA的PBS预包被30min后,洗涤,加入4μl DR4或DR5抗体,孵育1h后,洗涤,加入FITC标记二抗孵育1h,PBS洗涤2次后,加入1μg/ml PI后,流式细胞仪检测细胞FITC的荧光强度。
结果:TRAIL死亡受体是决定肿瘤细胞对TRAIL是否有敏感性的重要因素,本研究发现哇巴因能显著性提高A549细胞内DR4与DR5mRNA的表达量(图6)。对细胞膜DR4与DR5包被特异性抗体及FITC标记的二抗后,流式细胞仪技术分析细胞膜DR4与DR5的表达情况(图7),结果显示哇巴因能显著性增加细胞膜表面DR4与DR5的分布。
实施例六哇巴因联用TRAIL用药的体内抗肿瘤作用取指数生长期的A549细胞,经胰酶消化后,用PBS洗2遍。将每只100μL107个A549细胞皮下注射到7周龄的裸鼠右侧背部皮下,一周后形成原位肿瘤,将小鼠分成4组,每组8只。然后开始治疗,分别用PBS、哇巴因2μg/只、TRAIL100μg/只、哇巴因/TRAIL联合治疗组进行裸鼠腹腔注射,每天注射一次,共治疗15次。从开始治疗后每三天测量肿瘤大小,并按下列公式计算肿瘤体积:0.5236×L1×(L2)2,其中L1为肿瘤长轴,L2为肿瘤短轴。开始治疗后第25天,处死小鼠,进行数据分析。结果:接种A549细胞于裸鼠形成人源肺癌实体瘤,由于正常情况下,A549低表达TRAIL受体DR4与DR5,对TRAIL诱导凋亡作用具有很强的抵抗性,所以单用TRAIL抗肿瘤效果并不显著。而通过TRAIL与哇巴因联合治疗异体接种A549负瘤裸鼠,显示出明显的治疗效果,肿瘤体积显著小于其他三组(图8,9)。
Claims (1)
1. 哇巴因与肿瘤坏死因子相关的凋亡诱导配体TRAIL联用在制备治疗非小细胞肺癌药物中的应用。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN2010102355289A CN101869574B (zh) | 2010-07-24 | 2010-07-24 | 哇巴因在增强非小细胞肺癌细胞敏感性中的应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN2010102355289A CN101869574B (zh) | 2010-07-24 | 2010-07-24 | 哇巴因在增强非小细胞肺癌细胞敏感性中的应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN101869574A CN101869574A (zh) | 2010-10-27 |
CN101869574B true CN101869574B (zh) | 2011-12-21 |
Family
ID=42994780
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN2010102355289A Expired - Fee Related CN101869574B (zh) | 2010-07-24 | 2010-07-24 | 哇巴因在增强非小细胞肺癌细胞敏感性中的应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN101869574B (zh) |
Families Citing this family (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103536925B (zh) * | 2013-10-28 | 2015-07-01 | 中国医学科学院基础医学研究所 | 强心苷化合物在非小细胞肺癌治疗中的应用 |
CN103638036B (zh) * | 2013-11-22 | 2015-03-04 | 南京大学 | 强心甾类化合物在制备治疗脓毒症免疫麻痹药物中的用途 |
TWI701030B (zh) | 2015-09-25 | 2020-08-11 | 昇捷生物科技股份有限公司 | Ev71感染的組合治療 |
CN111166758A (zh) * | 2019-12-10 | 2020-05-19 | 安徽瀚海博兴生物技术有限公司 | 一种利用哇巴因的组合抗癌药物 |
CN110960543A (zh) * | 2019-12-10 | 2020-04-07 | 安徽瀚海博兴生物技术有限公司 | 一种联合抗vegf-抗pd1双特异性抗体用于制备抗癌药物 |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2000045165A1 (en) * | 1999-02-01 | 2000-08-03 | Cytovia, Inc. | Methods of identifying therapeutically effective antineoplastic agents with cultured cells having intact cell membranes and corresponding products |
WO2005060951A2 (en) * | 2003-12-19 | 2005-07-07 | Bionaut Pharmaceuticals, Inc. | Anti-neoplastic agents, combination therapies and related methods |
WO2006029020A2 (en) * | 2004-09-02 | 2006-03-16 | Bionaut Pharmaceuticals, Inc. | Treatments of refractory cancers using na+/k+-atpase inhibitors |
AU2008321225A1 (en) * | 2007-11-13 | 2009-05-22 | Phoenix Biotechnology Inc. | Method of determining the probability of a therapeutic response in cancer chemotherapy with cardiac glycoside |
Family Cites Families (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2721817B2 (ja) * | 1995-05-15 | 1998-03-04 | 萩原 義秀 | ヒト/ヒト・ハイブリドーマ及びその生産する抗体 |
JPH08308571A (ja) * | 1995-05-18 | 1996-11-26 | Asahi Chem Ind Co Ltd | 融合細胞の安定化剤 |
-
2010
- 2010-07-24 CN CN2010102355289A patent/CN101869574B/zh not_active Expired - Fee Related
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2000045165A1 (en) * | 1999-02-01 | 2000-08-03 | Cytovia, Inc. | Methods of identifying therapeutically effective antineoplastic agents with cultured cells having intact cell membranes and corresponding products |
WO2005060951A2 (en) * | 2003-12-19 | 2005-07-07 | Bionaut Pharmaceuticals, Inc. | Anti-neoplastic agents, combination therapies and related methods |
WO2006029020A2 (en) * | 2004-09-02 | 2006-03-16 | Bionaut Pharmaceuticals, Inc. | Treatments of refractory cancers using na+/k+-atpase inhibitors |
AU2008321225A1 (en) * | 2007-11-13 | 2009-05-22 | Phoenix Biotechnology Inc. | Method of determining the probability of a therapeutic response in cancer chemotherapy with cardiac glycoside |
Also Published As
Publication number | Publication date |
---|---|
CN101869574A (zh) | 2010-10-27 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Tan et al. | IL-17A plays a critical role in the pathogenesis of liver fibrosis through hepatic stellate cell activation | |
CN101869574B (zh) | 哇巴因在增强非小细胞肺癌细胞敏感性中的应用 | |
Chen et al. | p8 attenuates the apoptosis induced by dihydroartemisinin in cancer cells through promoting autophagy | |
CN102321158A (zh) | 阻止细胞dna合成抑制细胞增殖的多肽及用途 | |
CN104398526B (zh) | 雷公藤甲素和雷公藤红素在制备抗肿瘤药物中的应用 | |
CN101904837B (zh) | 鱼藤酮与trail联用在制备治疗非小细胞肺癌药物中的应用 | |
Jin et al. | Indocyanine green-parthenolide thermosensitive liposome combination treatment for triple-negative breast cancer | |
Zhou et al. | Calycosin enhances some chemotherapeutic drugs inhibition of Akt signaling pathway in gastric cells | |
Bespalov et al. | Experimental study of antitumour activity and effects on leukocyte count of intraperitoneal administration and hyperthermic intraperitoneal chemoperfusion (HIPEC) with dioxadet in a rat model of ovarian cancer | |
Li et al. | Artesunate possesses anti-leukemia properties that can be enhanced by arsenic trioxide | |
Vistica et al. | Therapeutic vulnerability of an in vivo model of alveolar soft part sarcoma (ASPS) to antiangiogenic therapy | |
CN104840482A (zh) | 一种中药有效组分组合物及其用途 | |
Wang et al. | Increased insulin-like growth factor 1 receptor (IGF1R) expression in small cell lung cancer and the effect of inhibition of IGF1R expression by RNAi on growth of human small cell lung cancer NCI-H446 cell | |
Ma et al. | Antineoplastic agents in chemotherapy facilitating tumor growth and angiogenesis in the interval administrations | |
CN106265760B (zh) | 戈氏梭菌驯化株在制备放疗增敏剂中的应用 | |
Lee et al. | In vitro and in vivo anticancer effects of destruxin B on human colorectal cancer | |
CN102499942A (zh) | 雷公藤甲素顺铂联合运用在制备抗耐药性胰腺癌药物中的应用 | |
CN101732329A (zh) | 一种具有协同作用的治疗t细胞淋巴瘤的组合药物及其应用 | |
CN101879215B (zh) | 五味子木脂素类成分在制备抗肺癌的药物中的应用 | |
CN106955292B (zh) | 一种治疗食管癌的药物组合物及用途 | |
CN102008715B (zh) | 抗肿瘤的MA-TNFα药物组合物及其应用 | |
Nakagiri et al. | Thymoma-associated graft-versus-host disease-like erythroderma | |
CN107446024A (zh) | 一种可拮抗ddx3蛋白rna结合活性的多肽dip‑13及其应用 | |
CN104083368A (zh) | G-1在制备基于g蛋白偶联受体30的三阴性乳腺癌靶向药物方面的应用 | |
CN107299138A (zh) | Cxcl4单抗治疗肿瘤及化疗后肿瘤加速再增殖基因筛选法 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
C14 | Grant of patent or utility model | ||
GR01 | Patent grant | ||
C17 | Cessation of patent right | ||
CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20111221 Termination date: 20130724 |