| Forewords:Benign prostate hyperplasia (BPH) is called as hyperplasia of prostate for short, which is a common disease of the older men. In china, as the extension of life expectancy, the improvement of life quality and living environment change, the prevalence is close to the developed countries. Although there is an upward trend in the incidence of hyperplasia of prostate, but its pathogenesis is yet to be clear. Although surgical treatment is the ideal method for treatment of prostatic hyperplasia, but the patients with the disease are for older people, so quite a few patients can’t tolerate surgery. If we can analyze its pathogenesis from the gene level, find the key sites and targeted therapy of the disease, to treat this part of the patients and reduce the social burden has a significant and positive significance.For research into the causes of prostatic hyperplasia, from the initial of cell morphology, and then deep into molecular field, different research methods and the establishment of the experimental model with hyperplasia of prostate genes and cytokines being discovered. The research direction mainly focus on four aspects of proliferation, differentiation, apoptosis, and senescence, in theory, have proposed the male estrogen imbalance, embryo to reawaken and polypeptide growth factor effect, etc. General speaking, everyone body’s tissues and organs will with the aging to degenerate, shown the degradation of cell proliferation ability, tissues and organs volume contraction. But the prostate, on the contrary, as the growth of the age showed the tendency of cell proliferation and increase of volume. To this end, we put forward the cell aging escape mechanism, that is the cells survive longer through to the lack of sensitivity on apoptosis on the one hand, and have more numbers by slowing down aging on the other hand. Its overall effect led to the increase of prostate cells. From the perspective of the histologic findings of BPH, this is a reasonable explanation to the pathogenesis of prostatic hyperplasia. Under the guidance of this idea, we first studied the importance of telomerase in this mechanism, At normal conditions, the telomerase activity in cells is tightly controlled, increased telomerase activity can lengthen cells telomeres, which will increases the number of cell proliferation and prolong life of cells. By detecting telomerase activity expression in prostate hyperplasia and normal tissue respectively, we found that the telomerase activity in hyperplasia tissues was higher than normal tissue, preliminarily obtained the aging escape exists in the hyperplasia of prostate. Further study we compared the expression of pl6iak4a (an representative aging gene) in the tissue of prostate hyperplasia and normal tissue, Found the expression of pl6iak4a genes is low in hyperplasia group, the low expression may delay the ageing of the cells, make the prostate cells escape age, lead to cells accumulation and tissue proliferation. All of above strongly affirmed the importance of aging escape mechanisms in the pathogenesis of prostate.The Bcl-xL is a protein of the Bcl-2family, play an important role in cell apoptosis regulation. The Bcl-xL with other romote apoptosis protein can form to heterodimer to inhibit apoptosis, also can format oligomers channel directly on the mitochondrial outer membrane to maintain mitochondrial membrane’s normal state during cellular stress. A large number of studies have shown that the antiapoptotic protein Bcl-xL in aging cells can make the cells has more resistance to apoptosis, thus destroy normal cells apoptosis, prolonged cellular survival time and to cumulation. Prostate hyperplasia as a typical age-related diseases, the relationship with Bcl-xL yet is not clear.It is Known during the growth and development of the prostate, androgens and its receptor play an important role, a large number of studies have shown that the expression of androgen receptors in the hyperplasia of prostate tissue is more than normal tissue significantly. So in here, a correlation analysis of expression of androgen receptors and BCL-XL in the hyperplasia prostate tissue is helpful to understand their interactions in the pathogenesis, provide the basis for further revealing the etiopathogenesis. And for the development of prostate hyperplasia’s molecular targeted therapy drugs provide important theoretical basis. Objective:In this experiment, we plan by collecting the hyperplasia of prostate tissue and normal prostate tissue, detecting the expression of antiapoptotic proteins BCL XL and androgen receptor in the two groups;Respectively analysis the relationship of antiapoptotic proteins BCL-XL and androgen receptors’ expression, the relationship between antiapoptotic proteins BCL-XL and prostate hyperplasia disease’s progress factors, provide theoretical basis for clinical targeted drugs treatment of prostate hyperplasia. Methods:1. Collectiing tissue samples of normal prostate and hyperplasia prostate. Normal prostate specimens come from the renal transplant donor with brain death, hyperplasia prostate tissue derived from the patients with radical prostatectomy or Bladder cancer radical. tissues are fixed48hours with4% formaldehyde after sampling. First make a HE staining after pathological section, Under microscope to identify the specimens as normal or hyperplasia tissue, then using paraffin embedding and seal. At the same time record the source of each specimen, date, information classification and related clinical indicators. The clinical risk progressivity indicators of Prostatic hyperplasia have age (years old), Prostatic volume (cm after), prostate specific antigen PSA (ng/ml), IPSS score (points) and the maximum urine flow rate Qmax (ml/s). The paraffin blocks of embedding specimens stored at low temperature environment of4℃, to prevent the specimen is damaged.2. Detection the expression of antiapoptotic proteins BCL-XL and androgen receptors. Paraffin blocks was made from tissue slides after biopsy. In turn by the baking sheets, dimethyl benzene dewaxing, gradient alcohol hydration to water, eliminate endogenous peroxidase with3%hydrogen peroxide solution, antigen high temperature repair use0.01mol/L pH6.0citrate buffer in the microwave oven, then make immunohistochemical staining:In0.01mol/L pH7.2～7.4PBS wash three times, every time is3min; add the first antibody and Placed in the4℃wet box for one night; then in turn with sealing, add the second antibody and the SABC reagent, after those, make DAB staining under microscope’s control, redyeing with Hematoxylin. At last to dehydration, transparent sealing slide and examination under microscope. In the experiment, use PBS take place of the first antibody in negative control group, positive control group using breast and lung cancer biopsy make a contrast. Image analysis:all the immunohistochemical slices are image acquisition and analysis with the Image pro-plus Image collection and analysis system under the same conditions. After20minutes Image acquisition system preheating, adjust and fixed the set, such as light source, the strength of aperture, white balance, exposure, sensitivity and contrast, and so on. to select representative area from each slice, according to the3x3intake9high power field of vision. Then import into the image analysis system to calculate integral optical density value (IOD) of each slice. Use IOD values divided by the measurement area, get the average optical density value (MD) as the result of a slice. Respectively to detect the expression of antiapoptotic proteins BCL-XL and adrogen receptors in the prostate hyperplasia tissues and normal prostate tissues, record the data of two grounps.3. Statistical analysis of experimental data. Comprehensive analysis the clinical indexes and the expression of antiapoptotic proteins BCL-XL and androgen receptor. The experimental data is expressed as (x±s), using SPSS13.0data packets for analysis. The correlation between two variables analysised by Spearman rank correlation. Take P<0.05for statistically significant differences. Respectively to assess the expression relationship between resistance apoptosis protein BCL-XL and androgen receptor, relations with prostatic hyperplasia (BPH) and the relationship with progressivity risk factors of prostate hyperplasia. Results:1. The expression and distribution of antiapoptotic proteins Bcl-x1and androgen receptor (AR) in prostate tissue. Immunohistochemical detection results show that: In normal prostate tissue, both epithelial and mesenchymal cells, the developing of Antiapoptotic proteins Bcl-x1are weak, prove its expression is very low. But the androgen receptor (AR) in epithelial cells and stromal cells have different degree of expression. It’s coloration is positive, and in the epithelial cells express more than mesenchymal cells. The coloration of antiapoptotic proteins Bcl-x1and androgen receptor (AR) in the hyperplasia prostate tissue are strong positive, explained the expression were significantly, and epithelial cells express more than mesenchymal cells. Analysis found that, the expression of antiapoptotic proteins Bcl-x1and androgen receptor (AR) more in hyperplasia prostate tissue than normal prostate tissue (P<0.05). And the rank correlation analysis showed that the expression of antiapoptotic proteins Bcl-x1and the expression of androgen receptor (AR) were positively correlated. Their correlation coefficient in normal group and BPH group is0.0399(P=0.1472) and0.5397(P=0.0028). Suggests that both the expression have a significant relative in benign prostatic hyperplasia tissues.2. The relationship of the expression of antiapoptotic proteins Bcl-x1and clinical indicators in patients with benign prostatic hyperplasia (BPH). In this study,30cases of benign prostatic hyperplasia (BPH) patients’ clinical testing indicators record as follows:Age is (67.3±6.9) years, prostate volume is (170.1±70.7) cm3, prostate specific antigen (PSA) is (3.7±6.1) ng/ml, IPSS score is (24.4±4.3), maximum urinary flow rate (Qmax) is (11.0±7.0) ml/s. After the Spearman rank correlation analysis shows that: The expression of Bcl-x1was positively correlated with patients’ age (P<0.05); and positively correlated with prostate volume (P<0.05); and positively correlated with PSA (P<0.05); but no significant correlation with IPSS score (P>0.05); and has no obvious correlation with maximum urinary flow rate (Qmax)(P>0.05). Conclusions:This study proves that the antiapoptotic proteins Bcl-xl is high expression in hyperplasia prostate tissue cells and have a positive correlation with androgen receptors. That strongly supported the speculation that antiapoptotic proteins Bcl-xl through synergy with androgen may cause aging prostate cells escape, causing the proliferation and accumulation of cells. In addition, analysis found that the Bcl-x1has a positive correlation with patients age, prostate volume and PSA value. Shows it is also promote the development of prostate hyperplasia to a certain extent. For prostatic hyperplasia’s development mechanism provides a new thought, and for a possible new targets drugs on the treatment provide a theoretical basis. |
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