| It is known that ovarian carcinoma is the number one cause of death from gynecologic malignancy. Most patients have extensively metastasized to intra-peritoneum before being diagnosed for the reason of no striking clinical syndrome during early stage of ovarian cancer development. Although worldwide gynecologists have invested lots of time and efforts in solving this touchy disease, the five-year survival rate of ovarian cancer still fluctuate around 30%. The studies of ovarian cancer show that cisplatin is the primary choice medicine. It can be effectively used to treat advanced and recurrent ovarian cancer during the past few years. Chemotherapy is currently still a most useful method of ovarian cancer therapy. The cisplatin is, as we know, a dose-dependent medicine. Although the more dose we administrate, the more effect we can achieve, normally, severe renal failure, ear and neural toxicity are incurred by large dose of cisplatin, which results in application of lower dose and unsatisfied therapy effect. With the advance of thermobiology in recent years, thermotherapy has become the fifth effective method of ovarian cancer after operation, chemotherapy, radiotherapy and immunotherapy. It is noteworthy that the associate application of thermochemotherapy to ovarian cancer seems to be more and more practical and effective at present.Objective The purpose of this experiment is directed towards testifying the coordinate function of thermochemotherapy and investigating the effects of ovarian thermal degree, time and medicine concentration on cytotoxic effect of cisplatin. The results will recommend optimized method to clinical combined treatment of ovarian cancer.Method The subject is human papilla serous cystadeno carcinoma of ovarian (SKOV3). These cells were used in experiments when cultivated for 4 days. (1)The cell was divided into 4 groups. Each of them were exposed in turn to different thermoldegree 37℃, 40℃, 43℃ and 45℃ respectively for 30 minutes (min), and were separated into 5 subgroups, which then were treated by different medicine concentration, 0ug/ml, 0.lug/ml, lug/ml, 10ug/ml,100ug/ml, respectively. Ultimately, MTT was used to detect the cytotoxic index of each subgroup. (2)The cells were divided into 5 groups and each group was exposed to 43 ℃ for 0 min, 7.5 min, 15 min, 30 min and 60 min, respectively. Each of the 5 groups was separated into 5 subgroups, which contained different medicine concentration, 0ug/ml, 0.1ug/ml, lug/ml, 10ug/ml, 100ug/ml. MTT was also used to detect the cytotoxicity index. (3)The coordinate effect of thermo-chemotherapy on cytotoxic effect in SKOV3 is optimal compared to the separation of thermotherapy and chemotherapy. (4) The cell apoptosis was detected by flow cytometry (FCM), transmission electron microscopy (TEM) and electrophoresis.Results (1)Cytotoxicity index elevated with the increase of medicine concentration. CI value elevated with the increase of temperature under the same medicine concentration. There was significant difference of CI between the temperature over 40 ℃ and 37℃. These results showed that the increasing temperature could enhance the chemotherapy effect of cisplatin under the same medicine concentration. (2)When the temperature and medicine concentration were unchanged, CI value was progressive with the heating time. There was significant difference between heating time >30 min and < 15 min. These results showed that it was effective only when heating time was over 30 min. (3)The effect of combining themotherapy and chemotherapy is optimal. (4)When heating time was over 30 min, under the same medicine concentration and heating time, the apoptosis rate increased with the progressive temperature by FCM detection. Typical apoptosis cell could be seen under TEM. A typical DNA ladder of apoptosis could be detected by molecular biology technique.Conclusion (1)The combined application of themotherapy and chemotherapy can coordinately kill cancer cell. When temperature is over 40℃ (<45℃) and heating time is over 30 min,...
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