ABSTRACT
This study examined the effect of silencing LRIG3 expression on the proliferation and apoptosis of bladder cancer T24 cells and explored the role of LRIG3 in the tumorigenesis of bladder cancer. Bladder cancer T24 cells were routinely cultured and pSilencer plasmids were employed to construct LRIG3 eukaryotic expression vector of LRIG3-siRNA, i.e., pSilencer-LRIG3-siRNA. After confirmation, the vector was transfected into HEK293 cells to make a replication-deficient adenovirus, pAd-LRIG3-siRNA, which was then introduced into bladder cancer T24 cells. RT-PCR, Western-blotting were performed to detect the levels of LRIG3 mRNA and proteins. Cells number was determined by using MTT test. Hoechst33258 staining, transmission microscopy, flow cytometery were conducted to examine the cell apoptosis. Three groups included a blank control group, a negative control group (containing non-interfering plasmids) and a pAd-LRIG3-siRNA group. Our results showed that the recombinant pAd-LRIG3-siRNA was successfully transfected into the bladder cancer T24 cells. The siRNA formed by the transcription of the recombinant plasmids resulted in significantly reduced expressions of LRIG3 gene and protein and significantly decreased cell proliferation and growth in the pAd-LRIG3-siRNA group as compared with the control group (P<0.01). The siRNA also caused apoptotic changes of some cells, with the apoptosis rate being (17.69±0.75)%, which was significantly different from that of the control group (P<0.01). It was concluded that recombinant pAd-LRIG3-siRNA plasmids could effectively decrease the expression of LRIG3 mRNA and proteins and, to some extent, inhibit the proliferation and promote the apoptosis of bladder cancer T24 cells. Silencing LRIG3 gene might be a novel alternative for the treatment of bladder cancer.
ABSTRACT
Objective By detecting the expression of caveolin-1 in osteosarcoma tissues,to explore its possible roles in the occurrence,progress and metastasis of osteosarcoma and its relationship with clinicopathological features.Methods The expression level of caveolin-1 protein in 48 osteosarcoma tissues and 24 normal bone tissues were detected by immunohistochemistry of SP method.Its relationship with clinical and pathological features were also analyzed.Results among the 48 osteosarcomas examined,38 were stained weak or negative for caveolin-1,whereas the strong caveolin-1 staining was observed in all the 24 normal bone tissues.The absorbent optical density value of caveolin-1 protein in osteosarcoma tissues and normal bone tissues were(0.2292 ± 0.0329) and(0.6428 ± 0.0028),respectively.The expression of caveolin-1 in osteosarcoma tissues was significantly lower than that in normal bone tissues (t =21.48,P ＜ 0.01 ).Caveolin-1 protein expression was significant correlated with the pulmonary metastasis of tumors ( t =11.84,P ＜ 0.05 ),but had no correlation with nge,sex,tumor size and pathologic grades.Conclusion The low expression of caveolin-1 may participate in osteosarcoma invasion and metastasis,and may be a new prognostic molecular marker for osteosarcoma and a new therapeutic target for tumor.
ABSTRACT
Objective To study the role of interleukin-6(IL-6)in the pathogenesis of prostate cancer and its clinical significance. Methods Immunohistochemistry and RT-PCR were used to detect the expression of IL-6 protein and mRNA in frozen prostatic adenocarcinoma,adjacent benign prostatic tissue,and prostate cancer cell lines PC-3 and LNCaP. The serum levels of IL-6 in patients with prostate cancer and healthy controls,and the supernatants of prostate cancer cell cultures were measured by using ELISA. Results The IL-6 protein levels in prostate cancer tissue and PC-3 cells were significantly higher than those in adjacent benign prostatic tissue and LNCaP cells. The serum IL-6 levels in the patients with prostate cancer were markedly higher than those in the healthy controls. The IL-6 levels in supernatants in PC-3 cells were notably higher than those in the LNCaP cells. Conclusion The IL-6 gene may act as an important regulator in prostate cancer progression and may be one of the causes of prostate cancer conversion from an initially androgen-dependent state into an androgen-independent state.
ABSTRACT
The association between the single nucleotide polymorphisms (SNPs) in -174G/C and -634C/G of interleukin-6 (IL-6) promoter region and prostate cancer was examined in the population of Han people in Hubei region. TaqMan PCR was employed for the gene-typing of -174G/C and -634C/G in promoter region of IL-6 gene to compare the prostate cancer patients and normal controls in terms of genotype frequency, allele frequency and risk of prostate cancer. Enzyme-linked immunosorbent assay (ELISA) was used for the detection of IL-6 concentration in peripheral blood of the patients with prostate cancer and the relationship between the IL-6 level and the genotype was studied. Our results showed that in all the subjects, the genotype of genetic locus -174G/C was found to be GG and no CG and CC were observed. There was a significant difference in gene frequency of GG, CG and CC of -634C/G and allele frequency of G and C between prostate cancer patients and normal controls (P<0.05) and the gene frequency of GG+CG increased with the clinical stages and pathological grades of prostate cancer. The IL-6 level in GG+CG group was significantly higher than that in CC group. It was concluded that no SNP in -174G/C IL-6 promoter region was found in the population of Han people in Hubei region. The SNP in -634C/G was, to some extent, associated with the development and progression of prostate cancer. The population with GG+CG genetype has higher risk for prostate cancer.