| Background and Objective:Prostatic carcinoma is one of the most frequent malignant tumors of males in many western countries.Although the incidence of prostatic carcinoma in China is obviously lower than that of European and American, in the past few years, with the improvement of population aging, living condition and test facility, the disease incidence has had an obviously ascendant tendency.Benign prostatic hyperplasia (BPH) is the most common benign disease of prostate in old males.Although previous studies revealed that prostatic carcinoma progression was a process involving multiple molecular alterations,the precise molecular mechanisms of prostate cancer are remain poorly understood. Both prostatic carcinoma and benign prostatic hyperplasia have involved overgrowth of the epithelial cells. The change of gene expression of BPH is normal,but the growth of the epithelial cells of prostatic carcinoma is characterized by accumulation of molecular abnormalities because of genomic instability.Abnormal gene expression is the fundamental-reason of prostatic carcinoma's development and evolutionary. Therefore, comparative analysis of gene expression in prostatic carcinoma and benign prostatic hyperplasia may provide important information which are related to malignant transformation of prostatic cells.In this study, we used the technique of mRNA differential display to screen differential express of genes in prostatic carcinoma and benign prostatic hyperplasia and to clone and identify it for finding new molecular Markers,which can use to the diagnose and therapy of prostatic carcinoma. Methods:First of all,we extracted total RNA from prostatic carcinoma tissues and benign prostatic hyperplasia tissues.Equivalent high qualitative RNA were used for fluorescent differential display(FDD) and reamplified by PCR. Twelve pairs of primers, which are combined by three anchored primers with fluorescence and four arbitrary primers were applied for the experiment.After PCR products were separated by polyacrylamide gel electrophoresis,the differentially expressed placental cDNA bands were identified through fluorescent scanning and excised from the gel and reamplified, then they were being cloned by linked with T-easy vector and transformated with EcoRâ… and Hindâ…¢. After the recombinant was filtrated, purified and identified, all cDNA fragments we obtained were sequenced. The cDNA sequence were analysed-for homologue in GenBank database by using BLAST software. Northern blot was performed for determining the expressing of cDNA fragments in two groups of tissues.Results:Eighty-seven differentially expressed cDNA bands were cut down from polyacrylamide gel,twenty of them were reamplified, cloned and sequenced. We have obtained fourteen cDNA fragments.In seven highly expressed fragments in prostatic carcinoma tissues,one has obviously homologous sequences and its function is known.Although four of them have homologous gene sequences, their functions are unknown. Two of them haven't the extremely homologous sequences.In seven lowly expressed fragments in prostatic carcinoma tissues,three of them have obviously homologous sequences and their functions are known.Although one has homologous gene sequences,its function is unknown. Two of them haven't the extremely homologous sequences.There is not any correlative information in one of them.Conclusions:1.There are obviously differences of gene expression between prostatic carcinoma tissues-and benign prostatic hyperplasia tissues. Fluorescent mRNA differential display can be used effectively find out differently expressed genes in prostate cancer.2. By means of clone, sequence and homologous analysis, we can identify different expressed gene fragments which are screened and further understand its function and important clinical significance in prostatic carcinoma.3. In this study, new different display gene fragments play an important significance to construct and consummate gene bank of prostatic carcinoma. |
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