42 1 Vol 42 No 1 2018 1 Journal of Jiangxi Normal University Natural Science Jan 2018 1000-5862 2018 01-0100-06 MCF-7 VEGF 1 2 1 1 1 1* 1 570228 2 70102 Mcf -7 Cck-8 TUNEL- DAPI Cck-8 Cck-8 TUNEL-DAPI 1 mg ml - 1 24 h Elisa VEGF VEGF 20 10 mg ml - 1 VEGF 5 2 1 mg ml - 1 VEGF Mcf-7 VEGF VEGF R 737 9 A DOI 10 16357 /j cnki issn1000-5862 2018 01 17 0 0 80 2 d Morinda citrifolia Linn noni 0 89 DMEM Cat No SH30243 01 HyClone TM 0 10 3-4 fetal bovine serum FBS life technologies TM 0 01 - CO 2 50 ml L - 1 CO 2 37 VEGF VEGFR2 1 2 200 mg VEGF 10 ml DMEM 30 min 1 000 r min - 1 3 min 0 22 μm millipore 20 mg ml - 1 VEGF 1 3 Cck-8 1 1 3 1 Cck-8 1 1 2 000 1 000 0 500 0 250 0 125 ml - 1 Mcf-7 96 100 μl 2017-08-06 31360066 Hys2016-34 1962- E-mail fwynoni@ 163 com
1 MCF-7 VEGF 101 6 1 spss 20 5 ~ 6 h Cck-8 6 h r 2 = 0 996 0 Cck-8 6 h 450 nm v Cck-8 v = 1 11 37 3 4 5 6 7 h 450 nm 3 A = A - A A r 2 Cck -8 Mcf-7 1 1 Cck-8 1 Cck-8 /h 3 4 5 6 7 r square 0 976 0 0 991 4 0 994 1 0 996 0 0 994 3 Sy x 0 010 430 0 008 010 0 008 191 0 008 395 0 011 860 1 3 2 1 4 TUNEL-DAPI 1 000 r 1 4 1 15 mm min - 1 5 min PBS ph 7 4 nest 120 g 200 ml 2 400 ml 3 ml - 1 100 μl 96 100 μl 15 s 3 20 10 5 2 1 mg 950 ml L - 1 3 ml - 1 5 6 3 1 4 2 TUNEL-DAPI 24 h 48 10 μl Cck-8 6 h PBS 2 10 ml - 1 450 nm A = A - 500 μl A / A - A 14-15 1 000 0 500 0 250 1 3 3 Cck-8 0 125 mg ml - 1 1 000 r min - 1 5 min TdT DNase 10 min PBS 2 3 ml - 1 4 40 ml 100 μl 96 24 h L - 1 PBS 15 minpbs 20 10 5 2 2 TUNEL 1 mg ml - 1 48 h 72 h TUNEL DAPI Cck-8 8 6 3 48 h 72 h TUNEL FTIC 10 μl Cck-8 6 h FTIC + DAPI 450 nm
102 2018 20 mg ml - 1 10 mg ml - 1 1 5 5 mg ml - 1 2 mg ml - 1 500 ml - 1 6 1 mg ml - 1 2 ml 24 h 48 h 20 mg ml - 1 10 mg ml - 1 1 000 0 500 0 250 0 125 mg ml - 1 5 mg ml - 1 2 mg ml 24 h - 1 1 mg ml - 1 1 000 r min - 1 4 5 min 72 h 20 mg ml 4 PBS - 1 10 mg ml - 1 2 PBS 50 ml - 1 1 ml 1 000 r min - 1 4 5 min 5 mg ml - 1 2 mg ml - 1 1 mg ml - 1 50 100 μl annixv-pi 1 h BD FTIC 2 2 Cck-8 PI 4 FTIC 2 2 1 Cck-8 2 PI 24 h Mcf-7 flowjo 7 6 1 6 Elisa VEGF 20 mg ml - 1 p < 0 01 10 mg ml - 1 5 mg ml - 1 p < 0 05 PBS 2 10 ml - 1 24 h IC 50 = 9 030 ± 0 956 mg 96 ml - 1 1 mg ml - 1 PBS 3 Mcf-7 24 h 2 0 1 0 0 5 0 2 0 1 mg ml - 1 14 600 μl 10 000 g 5 min 4 15 Eisa VEGF Elisa VEGF 1 7 spss 20 * p < 0 05 ** p < 0 01 p < 0 05 2 Mcf-7 24 h Cck-8 2 2 2 2 Cck-8 3 48 h 72 h Mcf-7 2 1 48 h IC 50 = 2 964 ± 0 430 mg ml -1 72 h IC 50 = 0 983 ± 0 007 mg ml - 1 20 mg ml - 1 10 mg 24 h ml - 1 5 mg ml - 1 48 h
1 MCF-7 VEGF 103 p < 0 01 5 mg ml - 1 2 mg ml - 1 1 mg ml - 1 72 h 48 h p < 0 05 2 tunel-dapi p < 0 05 / Mcf-7 mg ml - 1 /% p * p < 0 05 ** p < 0 01 Mcf-7 1 000 8 18 369 ± 0 040 0 012 0 500 8 17 445 ± 0 020 0 017 0 250 8 12 792 ± 0 060 0 021 0 125 8 10 400 ± 0 080 0 053 0 000 8 9 290 ± 0 032-2 4 5 flowjo 7 6 1 000 mg ml - 1 24 h 0 12 0 125 mg 3 mcf-7 48 h 72 h ml - 1 Cck-8 0 04 2 3 Tunel-dapi Mcf -7 24 h 4 ImageJ 2 5 VEGF 6 20 mg ml - 1 VEGF 10 5 2 1 mg ml - 1 VEGF VEGF 24 h 5 mg ml - 1 48 h 10 mg ml - 1 72 h 2 mg ml - 1 VEGF VEGF 3 Cck-8 4 mcf-7 24 h tunel-dapi Mcf-7 1 mg ml - 1 8 2 1 000 mg ml - 1 0500 mg ml -1 0 250 mg ml -1 0 125 mg 24 h DAPI-TUNEL ml - 1 1 000 0 500 0 250 0 125 mg ml 4 2 mg ml - 1
104 2018 5 * p <0 05 ** p <0 01 6 Mcf-7 24 h 48 h 72 h VEGF 10 mg ml - 1 VEGF 1 ~ 5 mg ml - 1 VEGF AnnixV-pi Damnacanthal 2- -3- -1- -9 10 VEGFR2 VEGF VEGF VEGF 2 VEGF VEGF Mcf-7 VEGF 4 Cck-8 Mcf-7 VEGF 1 < 5 mg ml - 1 J 2016 23 171-172 177 VEGF 2 Deng Ye Chin Young-won Chai Heebyung et al Anthraquinones with quinone reductase-inducing activity and < 5 mg ml - 1 24 h benzophenones from Morinda citrifolia noni roots J Journal of Natural Products 2007 70 12 49-52 3 Muhammad Abdul Aziz Abdul Omar Tamilselvan Subramani et al Damnacanthal is a potent inducer of apoptosis with anticancer activityby stimulating p53 and p21 genes in MCF-7 breast cancer cells J Oncology Letters VEGF 11 VEGF leptin 12 2014 7 5 1479-1484 4 Ta?kin Elif Ilkay Akgün-Dar Kadriye Kapucu Ay?egül et VEGF al Apoptosis-inducing effects of Morinda citrifolia L and doxorubicin on the Ehrlich ascites tumor in Balb-c mice J Cell Biochemistry & Function Online 2009 7
1 MCF-7 VEGF 105 2 96-97 11 5 J 2012 12 1593- D 2011 1598 6 12 Cladophora fascic- J 2011 15 3 290-291 ularis J 7 Javier A García-Vilas Almudena Pino- ngeles et al 2016 40 1 89-92 The noni anthraquinone damnacanthal is a multi-kinase 13 Leptin VEGF inhibitor with potent anti-angiogenic effects J Cancer Letters 2017 385 1-11 8 VEGF mrna J 2011 2 140-143 D 2014 10 D 2011 J 2014 22 1807-1811 14 VEGF survivin bcl-2 J 2005 6 525-529 9 VEGF 15 RG3 A549 VEGF J Survivin VEGF-c 2006 1 60-62 The Effects of Noni's Freeze-Dried Powder on Breast Cancer MCf-7 Cell Line and Endogenous VEGF by Tumor Cells QUE Yanpeng 1 SHENG Zhanwu 2 SHEN Jiaqi 1 ZHAO Shuai 1 XIA Dongnan 1 FU Wenying 1* 1 Institute of Life Science College of Tropical Agriculture and Forestry Hainan University Haikou Hainan 570228 China 2 Haikou Experimental Station Chinese Academy of Tropical Agricultural Science Haikou Hainan 570102 China Abstract Adding noni's freeze-dry powder into Incomplete medium to culture Mcf-7 cell This paper will compare cellular biological characteristics including Cck-8 flow cytometry TUNEL-DAPI staining and so on Cck-8 assay showed that the Cck-8 absorption value of the cells treated with aqueous solution of Noni fruit powder is lower than those with incomplete cultured cell light absorption value and it has negative correlation with time TUNEL-DAPI double staining showed that the apoptosis rate of the normal cell apoptosis is higher than the one with Noni powder aqueous solution treatment and the apoptosis rate was positively correlated with the concentration Flow cytometry results showed that the cells treated with Noni powder aqueous solution in the concentration of 1 mg ml - 1 would occurs early apoptosis within 24 h Enzyme linked immunosorbent assay Elisa method to detect endogenous VEGF content showed that the content of VEGF reduced after treated in aqueous solution with Noni powder of 20 mg ml - 1 and 10 mg ml - 1 but VEGF content increased after treated in aqueous solution with Noni powder of 5 mg ml - 1 2 mg ml - 1 and 1 mg ml - 1 concentration Noni powder solution inhibited the growth of Mcf-7 human breast cancer cell and the mechanism might be by combining suppressor cell with VEGF Key words noni Mcf-7 VEGF