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APOCB2006-06-001 Study on the Ultrafine Structure of Tumor Cell With npg Cell Research (2006) 16:S73-S106 S73npg © 2006 IBCB, SIBS, CAS All rights reserved 1001-0602/06 www.nature.com/cr APOCB2006-06-001 among the four different phases, in which we can see the Study on the ultrafine structure of tumor cell with atomic different character in the protrusion distribution and num- force microscopy ber. Particularly in the nanometer scale, there are some Jie Zhu, Zuodong Wang prominent domains like any field distribution; maybe, it's Department of Applied Physics, College of Science, Northwest the particular structure in the nano-space which related to A&F University, Yangling 712100, China the biologic magnetic or electromagnetic fields. According to the prominent distinguishment character in the AFM Multiple pathways are involved in maintaining the genetic image, we found the credible method to discriminate cells integrity of a cell after its exposure to physical environ- in different cycle or living condition with the AFM and the ments. Although repair mechanisms such as homologous proper techniques. recombination and non-homologous end-joining are Keywords: ultrafine structure; tumor cell; atomic force important mammalian responses to double-strand DNA microscopy; synchronization; cell cycle damage, cell cycle regulation is perhaps the most impor- Correspondence: Jie Zhu tant determinant of physical environments sensitivity. A E-mail: [email protected] common cellular response to DNA-damaging agents is the activation of cell cycle checkpoints. The DNA damage induced by physical environments initiates signals that can APOCB2006-06-002 ultimately activate either temporary checkpoints that permit The effect of solanine on DNA and RNA in the tumor cells time for genetic repair or irreversible growth arrest that Chenfeng Ji1, YuBin Ji1, Shiyong Gao1, Xiang Zou1, results in cell death. Such checkpoint activation constitutes Shenghui Wang2 an integrated response that involves sensor, transducer, 1Research center on Life science and environmental science of and effectors genes. The cell cycle phase also determines Harbin University of commerce, Harbin, China; 2Postdoctoral a cell’s relative environmental sensitivity, with cells being Programme, Institute of Materia Medica, Harbin University of most environmental sensitive in the G2-M phase, less sen- Commerce, Harbin, China sitive in the G1 phase, and least sensitive during the latter part of the S phase. But the structure always can show the Commonly called “nightshade”, Solanum nigrum L. is inner function, vice versa. Besides the method of inside- an annual plant belonging to the Solanaceae family. It to-outside, we can realize the detail of the tumor cell by is found throughout China, but is more abundant in the detecting the ultrafine surface structure. The advancement Northeast and North China regions. Its main contents are of scanning probe microscopy has introduced a powerful alkaloids, but it also contains vitamins and many trace method of detecting biological structural features. It is elements. There have been reports of the plant being used now generally accepted that contact mode atomic force in folk medicine to treat tumors with good effect. In ear- microscopy (AFM) does not lead to significant cellular lier experiment, we found that solanine can significantly damage under appropriate imaging conditions. The power prolong the survival time of H22 tumor-bearing mice. In of AFM methods in resolving the structure and dynamic the present experiment, we further observed the effect of operations of cellular systems is just being recognized. In solanine on the DNA and RNA in tumor cells of S180 and the latest tapping mode AFM, the cantilever is oscillated H22 mice. S180 and H22 mice were divided into solanine at high frequency; 3D topography of the surface can be (37.50, 18.75, and 9.37 mg/kg) groups, negative control generated under milder scanning conditions. For the other group, and Cytoxan (30 mg/kg) group, each of which was way, we can utilize the nanomanipulation of the AFM to given drug through sc. Levels of RNA and DNA in tumor see the interaction between inner molecules, and realize the cells in each group were measured using a laser scanning relationship of macromolecules. Aim: Through compared confocal microscope (LSCM). Confocal images obtained the different detecting condition, get the best method of all by scanning the sample with a laser beam not only reveal and found the proper parameters to describe the physical the distributions of DNA and RNA on different sections and and chemical properties in the AFM study on tumor cell changes in the RNA/DNA ratio, but can show intuitively in synchronal，thereby find the suitable experimental changes in DNA and RNA after the cell is damaged in a process an narration way. Results and Discussion：We state more clearly approaching the real life situation. The interdict the cell into four different cycles with colchicum results demonstrated in the 37.50 and 18.75 mg/kg solanine i.e. G1, G2, S and M phase. From the clear and beauti- groups, RNA/DNA ratio in the tumor cell of both S180 ful AFM topographies, we can easily get the differences and H22 mice was significantly lowered. According to APOCB Congress | http://www.apocb2006.org.cn npg Cancer Biology S74 results, we could concluded that Solanine could lower the Xiujuan Zhang1, Yubin Ji1, Shiyong Gao2 RNA/DNA ratio in the tumor cells of S180 and H22 mice, 1Postdoctoral Work Station of the Institute of Materia Medica, which may be one of the mechanisms for the antitumor Harbin University of Commerce, Harbin, China; 2Physiology effect of solanine. Section, Department of Pharmacy, China Pharmaceutical Uni- versity, Nanjing, Jiangsu , China Keywords: solanine; tumor cell; DNA/RNA Correspondence: Chenfeng Ji E-mail: [email protected] Research has shown that Sargassum fusiforme polysac- charide (SFPS), the effective element in Sargassum fusi- forme which is a brown alga of the Sargasaceae family, APOCB2006-06-003 has some inhibitory effects on tumor cells. In this study, Study on the apoptosis effect induced by isothiocyanates we further study the induction of the apoptosis of the hu- in broccoli on human gastric adenoma cells SGC-7901 and man gastric carcinoma cell SGC-7901 by SFPS and its 2+ 2+ its mechanism effect on intracelluar concentration of Ca ([Ca ]i), so as Xiang Zou1, Yubin Ji1, Lang Lang1, Wenjie Chi2, to uncover the mechanism for its anti-tumor effect. The Xiaodan Wu2 effect of Sargassum fusiforme polysaccharide on the cell 1Research center on Life science and environmental science of cycle and apoptosis of tumor cells is observed using the 2+ Harbin University of commerce, Harbin, China; 2Postdoctoral flow of cytometry (FCM). Intracellular Ca is marked 2+ Research of the Instiute of Materia Medica, Harbin University with Fluo-3/AM and [Ca ]i is measured by using laser of Commerce, Harbin, China scanner confocal microscopy (LSCM). We find that SFPS can inhibit the passage of the human gastric carcinoma cell To investigate the pro-apoptosis effect of isothiocyanates (SGC-7901) from the G0/G1 stage to the S stage of the cell (ITCS) on human gastric adenoma cells SGC-7901, and cycle and increase the index of apoptosis (APO %). The 2+ its mechanism, SGC-7901 cells were treated with different results show that before the treatment of SFPS, [Ca ]i in 2+ concentrations of ITCS. MTT assay was used to evaluate SGC-7901 cells is fairly constant. After treatment, [Ca ]i the influence of ITCS on cell proliferation. Flow cytom- rapidly rises before it declines, but rises again after CaCl2 etry was used to test ROS levels, intracellular mitochon- is added. Finally we can conclude SFPS can induce apop- 2+ drial transmembrane potential (Δψm), and hypodiploid tosis in tumor cells by increasing [Ca ]i in the tumor cells. 2+ 2+ apoptosis peak in SGC-7901 cells. The results showed The rise in [Ca ]i is because Ca ions are released from that ITCS obviously inhibited proliferation of SGC-7901 intracellular calcium stores. cells. When treated with 0, 15, 30, 60, 120, 240 μg/mL of Keywords: Sargassum fusiforme polysaccharide; SGC- ITCS for 24 h, ROS levels were (1.6±0.5)%, (2.0±0.3)%, 7901; apoptosis; anti-tumor activity (5.5±0.4)%, (25.8±1.4)%, (83.7±1.2)% and (97.4±4.2)%, Correspondence: Xujuan Zhang respectively; and Δψm were (98.6±4.3)%, (98.4±4.8)%, E-mail: [email protected] (95.7±5.4)%, (92.6±4.0)%, (74.0±5.6)% and (63.7±4.0)%, respectively; when treated with 0, 60, 120, 240 μg /mL of ITCS for 48 h, cell apoptotic rates were (4.3±1.6)%, APOCB2006-06-005 (9.1±3.8)%, (20.1±4.2)% and (55.4±4.9)%, respectively. Effects of solanine on Km and Vmax of NATase in HepG2 From the results above, we could draw the conclusion that cells ITCS generates ROS in gastric cancer SGC-7901 cells, Yubin Ji1, Shiyong Gao2, Xiang Zou1, Meixu Wan2, which causes mitochondrial membrane permeabilization Yujin Zhang2 and Δψm decrease, therefore, leads to apoptosis of SGC- 1Research center on Life science and environmental science of 7901 cells. Harbin University of commerce, Harbin, China; 2Postdoctal Research of the institute of Materia Medica of Harbin University Keywords: broccoli; isothiocyanates; apoptosis; reactive of commerce, Helongjiang Harbin, China oxygen speciesmitochondrial; mitochondrial transmem- brane potential To explore the effects of solanine on Km and Vmax of NA- Correspondence: Xiang Zou E-mail: [email protected] Tase of HepG2. METHODS: employing HPLC, using 2-AF as substrate, taking concentration of 2-AF as concentration of substrate, in intact HepG2 cells and their cytoplasm, APOCB2006-06-004 taking the speed of 2-AF being acetylated to 2-AFF by Study on the induction of tumor cell apoptosis by Sargassum NATase as the rate of NATase, using double reciprocal plot, fusiforme polysaccharide taking 1/S(the reciprocal of concentration of 2-AF) and Cell Research | www.cell-research.com npg S75 1/V(reaction rate of NATase) as coordinates, got regression APOCB2006-06-007 equation, calculated Km and Vmax.
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