The Study On Effects And Related Mechanism Of Energy Controllable Steep Pulse On Invasion And Metastasis Ability Of Human Ovarian Cancer Cells SKOV3

Steep pulsed electric fields can cause irreversible electrical breakdown on the tumor cell membrane and results in tumor cells death, it would become a new non-invasive or mini-invasive physical approach in the treatment of neoplasm. In our prophas research it were proved that the ECSP can effectly kill the tumor cell. But we discover that there were existing the residual tumour cell aroud the tumor organiz, when pulse electric field was weakened with the distance increasing. The problem that the Biology behavior of the residual tumour cell is whether or not to change and the invasion and metastasis ability of the residual tumour cell is whether or not to strengthen is very significance. In our studies we use the residual tumour cell treated by ECSP as experiment model in order to study it's biology behavior and change of the invasion and metastasis ability. PART ONE THE EFFECTS OF ENERGY CONTROLLABLE STEEP PULSE ON INVASION AND METASTASIS ABILITY OF HUMAN OVARIAN CANCER CELLS SKOV3Objective: To observe the effects and analysis the different parameter especial sublethal parameter of ECSP on invasion and metastasis ability of human ovarian cancer cells SKOV3.Methods: SKOV3 cells were divided into seven groups . Taking without lightning stroke group as negative control group. The positive control groups were treated by volchicine,5-fluorouracil, volchicine and 5-fluorouracil,; other groups were treated by three different parameter of steep pulse.Using mechanical method to determine cell adhesion ability,Using Transwell cave to determine cell in vitro invision and chemotaxis locomotory ability.Using inverted microscope to observe and zonatize counting cell population on tergal Transwell cave ;Using Haematoxylin and Eosin to dye the membrane of Transwell cave and using light microscope counting cell population on tergal Transwell caveResults: The invasion and movement phenomenon of the groups treated by colchicina combined 5-fluorouracil was completely vanish . The invasion and chemotaxis locomotory ability of the group treated by 5-fluorouracil was not obviously depress. The invasion and chemotaxis locomotory ability of the groups treated colchicines and three different parameter of ECSP was lower than that of negative control group (P<0.05Conclusion: The ECSP can inhibit the invasion and metastasis ability of SKOV3,and has direct correlation with dosage.PART TWOEXPERIMENTAL STUDY ON EFFECTS OF ENERGY CONTROLLABLE STEEP PULSES ON CYTOSKELETON OF HUMAN OVARIAN CANCER CELLS SKOV3Objective:The aim of our study was to determine the effects of energy controllable steep pulse (ECSP) on cytoskeleton of human ovarian cancer cells SKOV3 .Methods: SKOV3 cells were divided into five groups and treated by different parameter of steep pulse; Taking SKOV3 cells treated with volchicine as positive control group; and taking without lightning stroke group as negative control group. Using rhodamine-phalloidine to lable microfilament directly. Using immunofluorescence to label microbule and observing them by using Confocal Laser Scanning Microscope . Making specimen, and using electron microscope to observe the ultramicrostructure of cystoskeleton .Results : the immunofluorescence picture shows that SKOV3 cells whose energy controllable steep pulses treated lose the normal cystoskeleton network strcture. Obvious microfilament disaggregation, diffused skeleton protein , disappearance of cystoskeleton network stucture. Microbule disaggregation; reduction of pseudopodia; obvious microfilament disaggregation, permutation disorder and structrue disappearance.This effects has direct correlation with dosage.Conclusion: energy controllable steep pulse has obvious effect on SKOV3 cells' cystoskeleton structure . It can cause the disaggregation of microfilament and microbule, the reduction of cells'pseudopodia and the disappearance of network structure of cystoskeleton.PART THREE ELECTRIC PULSES REGULATE THE EXPRESSION OF MMP-9 AND TIMP-1 IN OVARIAN CANCERObjective: researchers have confirmed electric pulses have effects on cancer treatment. To further study the mechanism in cancer metastasis, the balance between matrix metalloproteinase (MMPs) and tissue inhibitor of metalloproteinase (TIMPs) are researched as the main reason of tumor metastasis.Methods: ovarian cancer bearing nude mice was subject investigated. And arranged randomly on experiment group and controlled group. Using electric pulses with voltage 1000V, frequency 1000Hz, duration 250ns treated on ovarian cancer tissue 5min, immunofluorscence was taken to detect the expression of MMP-9 and TIMP-1.Results: the fluorescence intensity of MMP-9 and TIMP-1 in controlled group were 140.21±5.16 and 74.98±3.14,the ratio was 1.87. in experiment group the fluorescence intensity respectively were 50.37±2.61 and 116.40±4.70, MMP-9 expressed lowly and TIMP-1 expressed heightened ,the difference was significant statistically(P<0.05), the ratio between MMP-9 and TIMP-1 was inverted to 0.43.Discussion: high intensity electric pulses regulated the balance of MMP-9 and TIMP-1, and indicated that the electric pulses inhibit ovarian cancer from metastasis and invasion, and also taken the evidence for clinical research of electric pulses.