Chinese Journal of Clinical Rehabilitation, Aug 2006, vol./is. 10/32(119-121), 1671-5926 (25 Aug 2006)
Wu Y.-G.,Ma Q.-J.,Dang G.-T.
Aim: To observe if the second filial generation of bisphosphonate-pamidronate disodium can inhibit the growth and induce the apoptosis of giant cell tumor (GCT) of bone cells. Methods: The experiment was finished from March 2004 to November 2005 in Third Hospital of Peking University. 1 The source of specimen: 10 fresh GCT tumor specimen was obtained from Department of Orthopaedics, Third Hospital, Peking University and Department of Orthopaedic Tumor Surgery of Beijing Jishuitan Hospital. All the specimens were confirmed by pathology. 2 The primary GCT cell was gained from the tumor tissue and cultured in vitro. After the tumor cells were adhered, the experiment groups were added with pamidronate disodium into culture dish, culture flask and 96 holes culture plate as medication group at 5, 25, 50, 100, 200 mumol/L, respectively. The blank control group was added with phosphate buffer saline (PBS). To observe if GCT cell growth was inhibited. 3 To detect the apoptosis of GCT cell with TUNEL: It was positive that there was amethyst particle in cell nucleus. The GCT cell apoptosis rate and the activation of caspase-3 were detected with flow cytometer. Results: 1 The culture of GCT cell: The cell of blank control group was normal and cell membrane was integrity. The cell of pamidronate disodium group was shrinked, pseudopodia were decreased, the contour was blurred, and the cell membrane was not integrity. The cell form was even disappear and disintegration. 2 The inhibition of GCT cell growth: Pamidronate disodium could inhibit the growth of GCT cell. Along with the increase of concentration and lasting of affecting time, it was observed that the inhibition of growth was more obviously. 3 TUNEL: The cell was found amethyst particle in cell nucleus of every concentration of pamidronate disedium group, while the blank control group was not found. 4 The apoptosis rate detected by flow cytometer: After treated with pamidronate disodium at 5, 50, 200 mumol/L for 48 hours, the cell apoptosis rate was obviously higher than that in the blank control group [(20.48+/-6.02)%, (42.39+/-12.41)%, (54.67+15.38)%, (10.13+/-3.45)%, P < 0.05]. 5 The detection findings of caspase-3 activation: After treated with pamidronate disedium at 5, 50, 200 mumol/L for 48 hours, the caspase-3 activation GCT cell was obviously higher than that in the blank control group [(14.93 +/-5.04)%, (25.13+/-7.60)%, (26.07+/-7.49)%, (2.73+/-0.81)%, P < 0.05]. Conclusion: Pamidronate disodium can inhibit the growth of GCT cell and induce GCT cell apoptosis by stimulating the activation of caspase-3. It was dose-dependent and time-dependent. It is indicated that the pamidronate disodium will be possible used in treatment and prevention of recurrence of GCT.