| BackgroundProstate cancer is the fifth most common cancer throughout the world. It is supposed t hat Asia enjoys a low incidence of prostate cancer, however, the incidence of prostate ca ncer there has been increasing year by year with the improvement of living conditions, t he changes in lifestyle and the progress in diagnosis. In China, the incidence of prostate cancer increases gradually especially in big cities. Hormone therapy is an effective treatme nt in th early stage of prostate cancer. Short or long term of hormone deprivation thera py can help to improve survival and delay the progress of the disease. However, after a medium period of 12 to 18 months, nearly all patients under the treatment of hormone deprivation therapies are progressing into the hormone independent stage of prostate ca ncer. Hormone therapy introduce no therapeutic actions in this stage, neither other thera pies. A rapid progression can be observed in this stage and most patients stride to the e nd of the life.Traditional Chinese medicine performs well in the clinical practice in treating different ki nds of cancers and it is more and more employed in cancer treatment these years for th e advantages of little side effect, multi-targets and multi functioning levels, focusing on c omprehensive regulation of the body compared to the modern therapies. The QAX presc ription used in our program is an effective QAX created by Prof. Pei Wang on the basi s of his experiences in treating cancers, which is called the QAX with the function of n ourishing the kidney and promoting blood circulation. The previous research in our labor atory indicated that this QAX should be used with cautions during the stage of androge n-dependent prostate cancer and it will give good performance on treating the disease du ring the androgen-independent stage. Besides, it can delay the disease progression from a ndrogen-dependent stage into the androgen-independent. LNCaP cells, the classic androge n-dependent prostate cancer cells, are used in our program to test the effects of the QA X above in vitro, so as to point out the proper moment to use the QAX with the func tions of nourishing the kidney and promoting the blood circulation in clinical practice.ObjectiveHuman prostate cancer LNCaP cells are employed in our program as the research model to observe the effects of the QAX with the function of nourishing the kidney and pro moting blood circulatioin on the biological behaviors of LNCaP cells in order to supply the research basis for expanding its clinical application. MethodsThe methods of intragastric administration of equivalent dose decoction and surgical rem oval of the testicles were employed to prepare 4 groups of different rat serums, the Nor mal Control group, the QAX group, the castration group and the Castration+QAX grou p. The serums described above are used to interfere the human androgen-dependent pros tate cancer LNCaP cells and MTT-asssav was used to measure the effects of the serums on cell proliferation; PI staining and PI/Annexin V double-staining were employed to te st the effects on cell cycle and apoptosis; Hoechst 34580 fluorescent staining was practic ed to observe the nuclear morphological changes of apoptosis cells caused by the serums; Fluo-4 AM fluorescent staining was performed to observe the time-lapse and instant cha nges of fluorescence indensity of cytosolic calcium caused by the serums with the help o f Laser Scanning Confocal Microscope; improved wounding assay was tried here to carry out continuous observation of the effects of the serums on the migration of LNCaP ce lls with the help of living cell station.Results1 Serums containing the QAX with the function of nourishing the kidney and promoting the blood circulation can accelerate the proliferation of LNCaP c ells, while as the castration group serum can suppress it. Compared to cast ration group, the Castration+QAX group can mildly accelerate the LNCaP prol iferation.2 Castration group serum can cause the G0/G1 cell cycle arrest in LNCaP cel ls. Compared to castration group, the Castration+QAX group can cause more si gnificant cycle arrest in the cells.3 After an interfereing period of 24 and 48 hrs, all groups performed signi ficant effects of promoting apoptosis in LNCaP cells aparting from the Norm al Control group, within which the castration group produced the strongest effect followed by the Castration+QAX group and at last the QAX group. The correspondingly typical nuclear morphological changes of apoptosis cells were observ ed by Laser Scanning Confocal Microscope.4 After 24 hrs of serums treatment, the cytosolic calcium indensity of LNCa P cells were observed to be increased. The castration group showed the high est indensity and then the QAX group and the Castration+QAX group the lowes t. After 48 hrs of serums treatment, the Castration+QAX group showed the hi ghest indensity followed by the castration group and the QAX group at last.5 The instant changing patterns of cytosolic calcium fluorescence indensity were different in different groups. The fluorescence indensity of calcium started to decrease in all groups exept the castration group.6 During the observing period of experiment, the migration speed of LNCaP c ells in Castration+QAX group was the fastest and then the castration group and the QAX group, the Normal Control group cells were the slowest, however, with no statistical significance.ConclusionThe QAX with the function of nourishing the kidnedy and promoting blood cir culation can mildly accelerate the proliferation of human prostate cancer L NCaP cells and can cause cell cycle arrest and apoptosis in the cells. Cast ration can significantly suppress the proliferation of the cells and cause cell cyecle arrest and accelerate cell apoptosis, which can be related to t he calcium overload. The QAX, castration or the combination of coumpound an d castration can accelerate the migration of LNCaP cells, however, consider ing the simple cultural conditions which can not simulate the enviroment in vivo, we should further our research on the effects on migration of the ce lls. |
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