| Objectives:To detect YKL-40 mRNA expression in eight kinds of tumor cells, study the effects of YKL-40 on proliferation, invasion and motility of prostate cancer LNcap cell, and investigate preliminarily the invasion mechanism.Methods:The expression of YKL-40 mRNA was detected by RT-PCR in eight kinds of tumor cell lines. The recombinant plasmid vector pcDNA3.1-YKL-40 was propagated, sequenced and transfected into prostate cancer cell line LNcap. The empty plasmid vector pcDNA3.1 transfection and non-transfection cells were used to be control groups. MTT method was used to determine the proliferation ability of the three groups of cells. Adhesion experiment and Boyden chamber assay was used to measure the changes of adhesion, invasion and motility activity. Semiquantitative RT-PCR was used to detect the mRNA levels of MMP-2, MMP-7, MMP-9, MT3-MMP, TIMP-1 and TIMP-2 in the three groups. Gelatin zymography was used to analyze the levels of MMP-2 and MMP-9 secretion and activities in the cells culture medium. Drug sensitivity test was used to examine the sensitivity to 5-fluorouracil, cisplatin and etoposide of LNcap cells before and after transfected by YKL-40.Results:Only prostate cancer cell line DU-145 and breast cancer cell line MCF-7 had YKL-40 gene expression in the eight kinds of tumor cells, and the expression level in DU-145 cell (1.100) was higher than that in MCF-7 cell (0.433). MTT assay showed that the OD49o of pcDNA3.1-YKL-40 transfected cells were significantly higher than the two control groups(P<0.05) at the second, third, fourth, fifth and sixth days. Adhesion experiment and Boyden chamber assay showed that the heterotypic adhesion(107.57%), invasion(75.11±4.40) and motility(133.00±5.07) of pcDNA3.1-YKL-40 transfected cells were increased significantly(P<0.05). However, the homotypic adhesion (1322±54,1562±79) was decreased(P<0.05). Semiquantitative RT-PCR results showed that compared with control groups, the mRNA expression of MMP-2, MMP-7 and MT3-MMP were raised in pcDNA3.1-YKL-40 transfected cells(the ratio of grayscale value:0.883±0.031,1.113±0.099,1.139±0.125, P<0.05). However, the TIMP-1 and TIMP-2 mRNA levels were reduced (the ratio of grayscale value:0.225±0.031.0.506±0.047, P<0.05). MMP-9 gene was not expressed in the three groups. Gelatin zymography confirmed that the secretion and activity of MMP-2 was heightened in the cells culture fluid of pcDNA3.1-YKL-40 transfected group, and there was no secretion of MMP-9 enzyme in the three groups of the cells culture fluid. In vitro drug sensitivity test demonstrated that the IC50 of 5-fluorouracil(31.15±0.43μmol/L), cisplatin(4.15±0.13μmol/L) and etoposide(55.22±0.57μmol/L) were also significantly higher than the two control groups(P<0.05).Conclusions:YKL-40 expression has variability in eight kinds of tumor cells. The proliferation, heterotypic adhesion, invasion and motility activities of LNcap cells are enhanced and the homotypic adhesion ability is depressed after YKL-40 transfection. The invasion mechanism may be by updating MMP-2, MMP-7, MT3-MMP mRNA expression, decreasing TIMP-1, TIMP-2 mRNA expression and heightening the secretion and activity of MMP-2 enzyme to degrade the extracellular matrix. YKL-40 also can decrease the drug-sensitivity of LNcap cells to 5-FU, cisplatin and etoposide. Therefore, YKL-40 may play an important role in the regulation of malignant transformation and local invasiveness of tumor cells. It may be a potential cancer therapeutic and prognostic molecular target. Objectives:To evaluate the diagnostic value of detection urine survivin mRNA with RT-PCR for bladder cancer by objective analysis on related studies with systematic review methods.Methods:With the search terms such as bladder neoplasm, survivin, RT-PCR, diagnosis, sensitivity, specificity, a systematic literature search was conducted in PubMed, SCI, EMBASE, Cochrane Library, CBM, CSJD, CMA digital periodicals, CJFD, Google Scholar and manual retrieval totally from 1997 to April 2009 to find all diagnostic trials with RT-PCR detections of urine survivin mRNA for bladder cancer. The Quality assessment of diagnostic accuracy studies (QUADAS) items and RevMan5.0 software were used to evaluate the quality and heterogeneity of the included studies. Fourfold table data was obtained from the included diagnostic test and used to calculate the summary values of sensitivity, specificity, positive likelihood, negative likelihood ratio by Meta-disc software. Summary receiver operator characteristic curve (SROC) was built and the area under SROC (AUC) was calculated also by Meta-disc software.Results:Twenty-six studies totally 2 416 met the eligibility criteria. Heterogeneity analysis showed that the heterogeneity among twenty-six studies was high(P<0.0001,I2=60%). Meta-analysis showed that compared with pathological examination, the summary values of sensitivity, specificity, positive likelihood ratio, negative likelihood ratio and AUC of urine-based survivin mRNA test using RT-PCR for bladder cancer were 88%,94%,14.56,0.13 and 0.9736. Sub-group analysis was conducted according to different RT-PCR technique. The results indicated that nested RT-PCR got the highest sensitivity, specificity and AUC and the values were 91%,95%, and 0.9805, respectively. The sensitivity and specificity of detection urine survivin mRNA with general RT-PCR were second, they were separately 87% and 94%. The sensitivity and specificity of using quantitative RT-PCR (QRT-PCR) was lowest in three sub-groups, they were separately 80% and 93%.Conclusions:The results of systematic review suggested that compared with pathological examination, the sensitivity and specificity of urine-based survivin mRNA test using RT-PCR is relatively high. It can be used as an important adjunct method for cystoscope in diagnosis and postoperative monitoring of bladder cancer.
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