Chinese Pharmacological Bulletin 2012 Sep ~ 6. http / /www. cnki. net /kcms /detail / R html

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1 ~ http / /www. cnki. net /kcms /detail / R html MG doi /j. issn A R R R R MG-63 MG Liu 2 Van Gieson Ⅰ 3 MG-63 D % ± 4. 69% G 0 /G 1 P < MG-63 MG-63 Ⅰ 1 MG cinnamic acid CINN MG-63 RNaseA PI DMSO Sigma RPMI 1640 Gibco 1. 2 MG-63 15% U L U No homewgh@ 126. com chifulee@ xmu. edu. cn L - 1 RPMI % CO 2 NaOH 4 3 mmol L h 3 mmol L - 1 ADM accumulation was much lower and mainly localized in the cytoplasm. From protein /DNA array 106 transcription factors among the total 345 transcription factors were up-regulated while 32 transcription factors were down-regulated. Conclusions The difference of transcription factors in MCF7 /W and MCF7 / ADM cells is probablely due to ADM s avoiding in entering the nuclei so that ADM cannot exert function. Key words MCF-7 /ADM adriamycin IC 50 transcription factor resistant cells extraction subcellular localization

2 osteonectin cells L ml D-Hank s ml 24 h 20 min 3 mmol L d x 珋 ± s 3 SPSS LSD d r min min2. 1 PBS ml PBS MG ml d MG-63 P < PBS 2 MG-63 DNase 100 mg L - 1 RNaseA min 50 mg L - 1 PBS 4 30 min % ± 4. 69% d 3 mmol L h HE Olympus BH D-Hank s r min min 2. 5% 4 1 h 1 mm % 4 1 h PBS 3 1% 4 2 h Fig JEM100-CX * P < Van Gieson Ⅰ 2. 2 Van Gieson MG-63 4 Bouin-Hollande Weigert G 0 /G 1 5 min P < S P < Van Gieson 5 min D-Hank s Bouin- Hollande min osteocalcin 7 d MG-63 P < Effect of cinnamic acid CINN on the proliferation of human osteosarcoma MG-63 cells ** P < vs control group MG-63 G 0 /G 1 Tab 1 Effect of cinnamic acid CINN on the cell cycle of MG-63 cells x 珋 ± s n = 3 Group G 0 /G 1 /% S /% G 2 /M /% Control ± ± ± 3. 1 CINN ± 1. 8 ** ± 1. 2 ** ± 2. 0 ** P < vs control group 2. 3

3 1264 MG-63 HE Fig 2A MG-63 5 HE Fig 2B 2. 4 G 0 /G 1 DNA MG-63 S MG-63 G 0 /G 1 S Fig 2C G 0 /G 1 MG-63 E G Fig 2D F H Ⅰ Van Gieson 7 Ⅰ MG-63 Fig 2I MG-63 Fig 2J I 2. 6 MG-63 Fig 2K 8 Ⅰ Fig 2L 90% 2. 7 Fig 2M Fig 2N 2. 8 MG MG-63 Ⅰ Fig 2O 5 Fig 2P 3 MG-63 Ⅰ osteosarcoma MG-63

4 1265 Fig 2 Changes in the morphology ultrastructure and the expressions of osteoblast markers of MG-63 cells before and after cinnamic acid CINN treatment A ~ B H. E staining C ~ H Ultrastructure under TEM N Nucleus ER Endoplasmic reticulum M Mitochondrion Calcified glycogen granules I ~ J Expression of typeⅠcollagen by Van Gieson staining K ~ L Bone nodules by alizarin red staining M ~ N Osteonectin expression by immunocytochemical staining O ~ P Osteocalcin expression by immunocytochemical staining MG-63 J. Mod J Int Tra- differentiation induced by single Chinese herb J dit Chin West Med Runx2 /Cbfal Osx product with potential use in cancer intervention J Runx2 /Cbfal Osx er J MG-63 of MGC-803 cells J Zhang R Cui Y A Zuo X D. Research progress on tumors cell 1. J Liu L Hudgins W R Shack S et al. Cinnamic acid A natural. Int J Canc-. 3 Lu J Wang H Lu F A. Effects of cinnamic acid on differentiation Chin Pharmacol Bull Van Gieson

5 Zhang S X Li H Chen D F. Exploration in collagen fiber staining method by Van Gieson J. J Prac Med Tech David M T Sandra A J Natalie A S et al. Terminal osteoblast differentiation mediated by runx2 and p27kip1 is disrupted in osteosarcoma J. J Cell Biol Birbeck M S C Symington T Carter R L M Lu X Chen J N Zhang J F. Raloxifene enhances osteoblastic differentiation in adipose derived stem cells via nitric oxide pathway Birbeck M S C Symington T Carter R L Translated by Yang J J. Chin Pharmacol Bull et al. The ultrastructure of tumour cells Basic science of tumour M. Beijing Science Press TPA skov3 11 Yang W Xu D H Zhou C H et al. Effect of berberine on the osteogenic J differentiation of bone marrow mesenchymal cells J. Chin Pharmacol Bull Gong P Li H X Zhang S M. The morphology and ultrastructure of human ovarian cancer cell line skov3 during differentiation induced by TPA J. Shandong Med J Wang Y P Ouyang Y M Jiang Y. Research progress of regulation of osteoblast differentiation and proliferation J. J Shanghai Jiaotong Univ Med Sci Aline B Latifa B Maria J et al. Fra-2 /AP-1 controls bone formation by regulating osteoblast differentiation and collagen production J. J Cell Biol J J Kristina K Catherine B K Anne M D et al. mir-29 suppression of osteonectin in osteoblasts Regulation during differentiation and by canonical Wnt signaling J. J Cell Biochem J Govoni K E Linares G R Chen S T et al. T-Box 3 negatively 8. regulates osteoblast differentiation by inhibiting expression of osterix and runx2 J. J Cell Biochem Effect of cinnamic acid on proliferation and differentiation of human osteosarcoma MG-63 cells WANG Guo-hong 1 GUO Zhi-yue 1 SHI Song-lin 2 LI Qi-fu 2 1. Dept of Physiology and Neurobiology Xinxiang Medical University Xinxiang Henan China 2. Dept of Basic Medicine Medical College Xiamen University Xiamen Fujian China Abstract Aim To investigate the effect of cinnamic acid on proliferation and differentiation of human osteosarcoma MG-63 cells. Methods MG-63 cells were treated with cinnamic acid. The living cells were counted by trypan blue staining. The morphology and ultrastructure of MG-63 cells were studied with optical microscope and electron microscope. The changes of cell cycle were analyzed by flow cytometry FCM. The expression of typeⅠcollagen was detected by Van Gieson staining. The formation of mineralized bone nodules was evaluated by alizarin red staining. The expressions of osteocalcin and osteonectin were detected by immunocytochemical staining. Results Cinnamic acid inhibited the proliferation of MG-63 cells significantly the inhibitory rate was % ± 4. 69% after treatment with cinnamic acid for 7 days. Flow cytometry a- nalysis showed that cinnamic acid increased the cell numbers in G 0 /G 1 phase significantly P < Under optical microscope and electron microscope the morphology and ultrastructure of MG-63 cells treated with cinnamic acid changed significantly compared with those in control group showing the characteristics of normal cells. Furthermore cinnamic acid promoted the expressions of osteoblast markers such as typeⅠcollagen osteonectin and osteocalcin increased calcium aggradation and facilitated the formation of bone nodules. Conclusion Cinnamic acid can inhibit the proliferation of human osteosarcoma MG-63 cells and induce the differentiation of MG-63 cells into osteoblasts. Key words cinnamic acid human osteosarcoma MG- 63 cells proliferation differentiation osteosarcoma osteoblasts