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10Figure 3-The 5-HT level of Drosophila brain changed in different time periods.(A) Different time periods female Drosophila brain 5-HT content have significant differences compared with the control group(*p<0.05,**p<0.01). (B)The fale flies have the same trend with the female flies.5-HT level displayed a significant compared with the control(*p<0.05,**p<0.01).
PAMD and its monomeric components on BxPC-3 Pancreas Tumor cells Proliferation Effect
Yang Li
HeilongjiangUniversity of Traditional Chinese Medicine,Haerbin,China
Abstracts:Objective:Explore PAMD, Dau, DS and proportion of group (Dau mixed 3:1 with DS ) drugs is proliferation of BxPC-3 pancreatic tumor cells in vitro. Method: BxPC-3 cells were subcultured, were added to low high school three doses PAMD, DS, Dau and proportion of group solution , while the control group an equal volume of saline, the role of 72h, by MTT assay absorbance (OD) calculate the group BxPC-3 cell proliferation inhibition rate. Result: The results showed that, PAMD high school low-dose group, the inhibition rates were 93.27%, 24.64%, 15.45%, Dau high school low-dose group, the inhibition rates were 67.18%, 24.09%, 12.00%, DS High School low dose inhibition rates were was 20.18%, 15.64%, 6.27 %, and the proportion of high school low -dose group, group inhibition rates were 30.18%, 41.27%, 17.36%. With the control group, in addition to the low-dose group of DS Dau groups were statistically significant (P <0.05). Conclusion: PAMD and its monomer component Dau and DS on BxPC-3 cell proliferation was inhibited, but Dau, DS and proportion group of drugs on BxPC-3 cell proliferation was weaker than PAMD group, indicating PAMD on BxPC-3 cells may also inhibit the proliferation of other ingredients with PAMD relationship.
Keywords:PAMD,Dauricine,BxPC-3, Proliferation
Chinese medicine is Menispermi Menispermaceous plants Menispermum dauricum (DC) of dry roots, while Phenolic alkaloids of Menispermum dauricum (PAMD) is extracted Menispermi soluble mixture of alkaloids[1]. PAMD, containing mainly Dauricine, (Dau) and Daurisoline,(DS), etc. Our previous studies have shown that, PAMD variety of tumor cells proliferation and
cytotoxicity [2'3], and its antitumor mechanism were studied[4-6]. On this basis, the experimental observation of different concentrations of PAMD, Dau, DS and proportion of group BxPC-3 cell proliferation, to further clarify the role of PAMD antitumor effective components, the following research.
1 . Materials and Methods
1.1 Materials
1.1.1 Cells and Reagents
BxPC-3 cell line by the Shanghai Fu Xiang Biotechnology Co., Ltd. to provide, modified RPMI-1640 medium, fetal bovine serum, trypsin and two resistance ( penicillin 100p,g/ml, streptomycin 100pg/ml) by Thermo Fisher biochemical products Co., Ltd. to provide, PBS by Beijing Zhongshan Golden Bridge Biotechnology Co., Ltd. to provide , MTT provided by the Sigma, USA , DMSO provided by the Amresco, other reagents were analytical grade .
1.1.2 Drugs
PAMD by the Heilongjiang University of Chinese Medicine College of Pharmacy Department of Traditional Chinese Medicine Wang Dong teachers, Dau by the Shanghai Biological Technology Limited, DS by the Shanghai Chemical Co., Ltd. by the Branch provided.
1.1.3 Instruments
Beijing East Hal Instrument Manufacturing Co., Ltd. produces clean bench (HDL), Germany HERAEUS company produces CO2 incubator (BB5060UV), provided by the Japanese Nikon inverted microscope (TS100), Nantong Science Instrument Factory desktop dried box (202-A0), Shanghai Shen An instrumentarija production of vertical pressure steam sterilizer (LDZX-75KBS), experimental equipment Co., Ltd. on the Hai Fuma electric oven (DPX-91628-2), Hunan Kay Limited production of scientific instruments centrifuge (TD6M), U.S. Thermo company provides ultra-low temperature freezer (994), East tube plant enzyme-linked immunosorbent assay (DG5031) and so on.
1.2 Methods
1.2.1 Cell culture
BxPC-3 cells in a 5% carbon dioxide incubator conditions 37 °C incubated with 10% fetal calf serum , penicillin (100pg/ml), streptomycin (100pg/ml) RMPI1640 subcultured in liquid culture, cells were used in experiments logarithmic growth phase.
1.2.2 The test product preparation
With the former are 1mol/l hydrochloric acid solution to dissolve, 1mol/l sodium hydroxide solution and adjusted to pH 6.0 , added to volume with saline and disposable filter, and the final concentrations were adjusted PAMD 80pg/ml, 40pg/ml, 20pg/ml, DS concentration of 16pg/ml, 8pg/ml, 4pg/ml, Dau concentration of 48pg/ml, 24pg/ml, 12pg/ml, the proportion group concentration of 64p,g/ml, 32p,g/ml, 16p,g/ml. Drug configured backup solution stored at 4 °C .
1.2.3 MTT determine BxPC-3 cell restrain disposed by Deugs
Logarithm growthcell cells adjusting the cell concentration of 1*106 /ml, inoculate in 96 orifice plate, 100^l per orifice,foster in 37°Cand 5% CO2 incubator.Incubation for 24h, were added to a concentration of 80,40,20 p,g/ml solution of PAMD concentration of 16,8,4 p,g/ml of DS solution, the concentration of 48,24,12 p,g/ml solution of Dau concentration is 64,32,16 p,g/ml solution of the proportion of groups , Each hole 20pl, blank control group plus saline 20pl, each group of six wells, and cultured 72h, before the end of the culture 4h, each culture well was added MTT (5mg/ml) 10pl, cultured 4h, the supernatant was discarded added to each well 10% DMS0100^l, concussion after 10min, using a microplate reader at 490nm wavelength detection of the optical density of each well (OD). The experiment was repeated three times. Collect receipts, calculation of drugs on the inhibition rate.
Inhibition ratio (%) = (1 - OD value of experimental group/control group OD value) * 100%.
1.2.4 Statistic test
Experimental data to x ±S, said the statistics with Excel software formula * , SD. Each drug concentration group compared with the control group t test was used, P <0.05 as the significance level .
2 . Results and discussion
2.1 Results:PAMD and its monomer composition on BXPC-3 cell line proliferation
MTT results showed that different concentrations of PAMD, DS, Dau and proportion group ( except low-dose group and DS Dau outside ) than the control group on BXPC-3 cells were proliferation , the difference was statistically significant (P <0.05), and the high-dose group PAMD BXPC-3 cells, inhibition rate reached 93.27% . Table 1 and Figure 1.
Table 1 PAMD and the monomer component on proliferation of BxPC-3 cells_
group Massconcentrati on (^g/ml) OD of BXPC-3 mean ( *±S, n=6) BXPC-3 inhibition ratio (%)
Blank control — 1.100±0.002 —
80 0.074±0.003*** 93.27
PAMD 40 0.829±0.001*** 24.64
20 0.930±0.001* 15.45
48 0.361±0.045*** 67.18
Dau 24 0.835±0.001** 24.09
12 0.968±0.014 12.00
16 0.878±0.001** 20.18
DS 8 0.928±0.003* 15.64
4 1.031±0.006 6.27
proportion 64 0.768±0.002*** 30.18
32 0.646±0.007** 41.27
group 16 0.909±0.001** 17.36
Note: Compared with the control group * P <0.05, ** P <0.01, *** P <0.001.
1.2
X±S,n=6
High dose Middle dose Low dose
I
PAMD
Dau
DS
Proportion Drug groups
Figure 1 PAMD and its monomer component in the proliferation of on BxPC-3 cells 2 . 2 Discussion
Pancreatic tumor is the most common malignant gastrointestinal tract, known as the "tumor of the King" , and its high degree of malignancy , rapid development, has discovered late, short course, early metastasis , poor prognosis [7]. In recent years, the incidence of pancreatic tumor showed an
upward trend at home and abroad, in China ranks the first six tumor mortality [8]. In the treatment of pancreatic tumor, in addition to the traditional surgical treatment, as well as chemotherapy, radiation therapy, targeted therapy and Chinese medicine treatment [9-13]. Application for comprehensive regulation of Chinese medicine therapy has become a hot research field in recent years .
This experiment clearly PAMD effective component of anti- pancreatic tumor and for further study. PAMD Dau and DS mainly contains two components, both of which accounted for about 80% of total PAMD. Which accounts for about 60% while the DS Dau approximately 20%, Dau with DS content ratio is about 3:1.Therefore, the composition of the experimental simulation PAMD grouped set different concentrations PAMD, Dau, DS and proportion group (Dau and DS by 3:1) the administration group, while a blank BxPC-3 cells cultured in the control group.Experimental results show that the high-dose group of drugs intensity of the effects were PAMD>Dau>proportion group>DS, namely the high-dose group PAMD BXPC-3 cell proliferation strongest possible with the other components PAMD on the antitumor effect of the two drugs may have promoted, or other component has some anti -tumor activity may also be PAMD multi-component, multi-target, multi-effects, synergies and overall adjustment related to the role; Each group of drugs in low dose group, the proportion of group inhibition than Dau strong group and the DS group, indicating that the two drugs may have a synergistic effect.PAMD on BxPC-3 cell proliferation inhibition of specific components and mechanisms for further study. References:
[1] Yunming Su,Yongqiang Li, Yuan Zhou. Bats phenolic alkaloids on experimental myocardial ischemia protective effect [J]. Chinese pharmacist , 2002, (06):326 -328 .
[2] Yunming Su, Caroling Xiao, Zhiguo Wang, etc. bat phenolic alkaloids on human tumor cells PC-3, BT6537 value of [J]. HarbinMedicalUniversity newspaper , 2007,41 (2):129 -131
[3] Yunming Su, Xing Meng, Xin Wang, et al. Bats phenolic alkaloids on K562, BXPC-3 Proliferation [J]. GansuCollege, 2008, (03) :1 -4.
[4] Yunming Su, Yihang Li, Kun Liu, etc. Bat phenolic alkaloids on BXPC-3 tumor tissue levels of MMP-2 [J]. ChengduMedicalCollege, 2010, (02) :113 -115 .
[5] Yunming Su, Kun Liu, Yan Zhang, et al. Bats phenolic alkaloids on BXPC-3 tumor inhibition and tumor tissue levels of MMP-9 [J]. Journal of Traditional Chinese Medicine , 2010, (01) : 33 -34 .
[6] Yunming Su, Yan CUI, Weijie Li, etc. Bat phenolic alkaloids on BxPC-3 tumor in nude mice and its effects on the expression of bFGF [J]. ChengduMedicalCollege , 2009, (04) : 245 - 247.
[7] Jieheng Wu, Mengna Liu, Shamariko HUANG, etc. pancreatic tumor animal model research progress [J]. JilinMedicalCollege , 2011,32 (2) :115- 117.
[8]Lopes RB,Gangeswaran R, McNeishIA,et al. Expression of the IAP protein family is dysregulated in pancreatic tumor cells and is important for resistance to chemotherapy [J]. Int.J.Tumor, 2009,120(11):2344-2352.
[9] Jinming Yu, Yong- hua YU, Yong YIN, et al . X -ray stereotactic radiotherapy analysis of 13 cases of pancreatic tumor [J]. Chinese Journal of Radiation Oncology , 2001,10 (2) : 125 .
[10] Shenyu Li, Suxian Wang, Xiangdong Cui, etc. pancreatic tumor new method - intraoperative brachytherapy radiotherapy [J]. Practical Oncology, 1991,5 (1): 4-6.
[11]Moore M J,Goldstein D,Hamm J,et al.Erlotinib plus gemcitabine compared with gemcitabine alone in patients with advanced pancreatic tumor: a phase III trial of the National Tumor Institute of Canada Clinical Trials Group[J].J Clin Oncol,2007,25(15) : 1960-1966.
[12]Troiani T,Martinelli E,Capasso A,et al.Targeting EGFR in pancreatic tumor treatment[J].Curr Tumor Drug Targets.2011,11(6):698-713.
[13]Matsuo Y,Takeyama H,Guha S,et ai.Cytokine network: new targeted therapy for pancreatic tumor[J].Curr Pharm Des.2012,18(17):2416-9.
