Expression of MiRNA-21 in diffuse large B cell lymphoma and its significance / 中国实验血液学杂志

MicroRNA-21 (miR-21) is considered to play a key role in many cellular processes, affecting tumorigenesis by inhibiting target gene expression. However, its role in diffuse large B-cell lymphoma (DLBCL) is still unclear, and there are no in depth studies on relationship between miR-21 and cellular phenotype. This study was aimed to investigated the expression and role of miR-21 in the regulation of cell biological behavior in DLBCL. The expressions of miR-21 in three DLBCL cell lines were detected by real-time quantitative reverse-transcription polymerase chain reaction (qRT-PCR). The possible roles of miR-21 in the biological and behavioral properties of DLBCL were explored by transfection of anti-miR-21 for miR-21 knockdown. In addition, PDCD4 and PTEN were assessed by luciferase reporter assay, qRT-PCR and Western blot. The results revealed that miR-21 expression was significantly upregulated in activated B-cell-like DLBCL cells as compared to germinal centre-like DLBCL cells. The inhibition of miR-21 could induce suppression of proliferation and invasion, as well as increase apoptosis in DLBCL. Moreover, knockdown of miR-21 increased the expressions of PDCD4 and PTEN at the protein level but not at the mRNA level. It is concluded the miR-21 can regulate proliferation, invasion, and apoptosis, so it has a potential therapeutic application in DLBCL.

Serum sCD44v6 and sE-cadherin levels in patients with esophageal squamous cell carcinoma / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the serum levels of sCD44v6 and sE-cadherin (sE-cad) in patients with esophageal squamous cell carcinoma.</p><p><b>METHODS</b>The serum samples were collected from 65 cases of esophageal squamous cell carcinoma, 32 cases of erosive esophagitis and 35 healthy subjects. Serum sCD44v6 and sE-cad levels were measured by enzyme linked immunosorbent assay (ELISA).</p><p><b>RESULTS</b>The mean levels of serum sCD44v6 and sE-cad in esophageal squamous cell carcinoma patients were significantly higher than those of erosive esophagitis patients and normal controls (both P<0.05). There was no significant difference in serum sCD44v6 and sE-cad levels between erosive esophagitis patients normal controls (P=0.566 and P=0.708, respectively). Serum sCD44v6 and sE-cad levels of esophageal cancer patients were not correlated with their clinicopathological features. Serum sCD44v6 level is not correlated with sE-cad level in squamous cell carcinoma patients(P=0.651).</p><p><b>CONCLUSION</b>Serum sCD44v6 and sE-cad might be a potential marker for screening of esophageal squamous cell carcinoma.</p>

Correlation between polymorphism in the promoter of DNA methyltransferase-3B and the risk of colorectal cancer / 中华预防医学杂志

<p><b>OBJECTIVE</b>To explore the correlation between the polymorphism in the DNA methyltransferase-3B (DNMT3B) gene promoter single nucleotide polymorphism (SNP)-149C→T (rs2424913) and-579G→T(rs1569686) and the genetic susceptibility to colorectal cancer in Jiangsu population.</p><p><b>METHODS</b>Genomic DNA was extracted from the leukocyte cell of blood samples collected from 544 colorectal cancer (CRC) patients (including 280 cases of colon cancer and 264 cases of rectal cancer) since January 2009 and July 2010, in a hospital, Jiangsu Province. The same samples were collected from the other 533 control subjects. Polymerase chain reaction with restriction fragment length polymorphism (PCR-RFLP) and DNA sequencing analysis were employed to assess the polymorphism of DNMT3B gene promoter-149C→T and-579G→T.</p><p><b>RESULTS</b>For DNMT3B-149C→T, no significant deviation was observed in the genotype distributions of polymorphisms between CRC cases (TT: 98.90% (538/544); CT: 1.10% (6/544)) and controls (TT: 97.75% (521/533); CT: 2.25% (12/533)) (χ(2) = 2.07, P = 0.15). The CC genotype was not detected in either patients or control subjects. The DNMT3B-149CT genotype was not associated with the risk of CRC (adjusted OR = 0.48, 95%CI: 0.18 - 1.30). For DNMT3B-579G→T, the genotype distributions of polymorphisms in CRC patients (TT: 90.07% (490/544); GT: 9.19% (50/544); GG: 0.74% (4/544)) were significantly different from those in control group (TT: 81.80% (436/533); GT: 17.82% (95/533); GG: 0.38% (2/533)) (χ(2) = 15.49, P < 0.05). The results showed that the-579 G allele could significantly decrease the risk of CRC (adjusted OR = 0.50, 95%CI: 0.35 - 0.72) in comparison with the -579 TT genotype. In addition, stratification analysis showed that for DNMT3B-579G→T, the genotype distributions of polymorphisms in colon cancer (TT: 92.50% (259/280); GT: 7.50% (21/280)) were significantly different from those in the controls (TT: 81.80% (436/533); GT: 17.82% (95/533); GG: 0.38% (2/533)) (χ(2) = 13.53, P < 0.05); and similar result was found in rectal cancer (TT: 87.50% (231/264); GT: 10.98% (29/264); GG: 1.52% (4/264)) and controls (TT: 81.80% (436/533); GT: 17.82% (95/533); GG: 0.38% (2/533)) (χ(2) = 5.64, P = 0.018). G allele carriers could decrease the risk of colon cancer (adjusted OR = 0.38, 95%CI: 0.23 - 0.63), and the risk of rectal cancer (adjusted OR = 0.65, 95%CI: 0.42 - 0.99). However, for DNMT3B-149C→T , there were no significant deviation in the genotype distributions of polymorphisms between colon cancer (TT: 98.57% (276/280); CT: 1.43% (4/280)) and controls (TT: 97.75% (521/533); CT: 2.25% (12/533)) (χ(2) = 0.82, P = 0.366); and there was no significant deviation between rectal cancer (TT: 99.24% (262/264); CT: 0.76% (2/264)) and controls (TT: 97.75% (521/533); CT: 2.25% (12/533)) either (χ(2) = 1.89, P = 0.169).</p><p><b>CONCLUSION</b>Our research demonstrates that the-579 G allele is a potential protective factor for the occurrence of CRC, however, the polymorphism of DNMT3B-149 gene shows no close correlation with the occurrence and development of CRC among Chinese population.</p>

Effect of recombinant adenovirus Ad-DT-A in targeted therapy for malignant cancer cell lines with loss of IGF2 imprinting / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To explore the feasibility of IGF2 imprinting system in target gene therapy for tumors.</p><p><b>METHODS</b>The mouse H19 enhancer, DMD and promoter H19 were amplified by PCR from mouse genomic DNA and then cloned into the plasmid pDC312. The EGFP and DT-A fragments were amplified by PCR and cloned into the recombinant plasmid, and then the shuttle plasmid were transfected into HEK293 cells together with the adenoviral vector Ad5, namely, Ad-EGFP and Ad-DT-A. Adenovirus hexon gene expression was applied to confirm the presence of adenovirus infections. The effect of the IGF2 imprinting system was tested by fluorescence microscopy. RT-PCR and Western blotting after transfection of the recombinant adenoviral vectors into cancer cells were used to show loss of IGF2 imprinting (LOI) and maintenance of IGF2 imprinting (MOI), respectively. The anti-tumor effect was assessed by MTT and flow cytometry after the HCT-8 (LOI). Human breast cancer cell line MCF-7 (MOI) and human normal gastric epithelial GES-1 (MOI) cell line were transfected with Ad-DT-A in vitro. The anti-tumor effect was detected by injecting the Ad-DT-A in nude mice carrying HCT-8 tumors.</p><p><b>RESULTS</b>The expression of EGFP protein, DT-A mRNA and DT-A protein were seen to be positive only in the HCT-8 tumor cell line. Infection with Ad-DT-A resulted in obviously growth inhibition in HCT-8 cells (75.4 ± 6.4)% compared with that in the control group, and increased the percentage of apoptosis in the HCT-8 cells (20.8 ± 5.9)%. The anti-tumor effect was further confirmed by injecting the recombinant adenoviruses in HCT-8 tumor-bearing nude mice, and the results showed that the Ad-DT-A inhibited the tumor growth, with on inhibition rate of 36.4%.</p><p><b>CONCLUSIONS</b>The recombinant adenoviruses carrying IGF2 imprinting system and DT-A gene have been successfully constructed, while Ad-DT-A can effectively kill the tumor cells showing loss of IGF2 imprinting. It might play an important role in future target gene therapy against malignant tumors based on loss of IGF2 imprinting.</p>

Study on the relationship between polymorphism sites of IL-10 and the susceptibility of endometriosis / 中华预防医学杂志

<p><b>OBJECTIVE</b>To explore the relationship of the endometriosis susceptibility and polymorphism of up stream of IL-10 promoter at the site of 1082(G→A), 819(C→T) and 592(C→A).</p><p><b>METHODS</b>A total of 214 patients with endometriosis and 160 healthy individuals were enrolled and divided into patient group and control group in this study. The polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) was applied to detect the base transition in the up stream of IL-10 promoter at the site of 1082(G→A), 819(C→T) and 592(C→A). SPSS11.0 software was applied to analysis frequencies of all genotypes.</p><p><b>RESULTS</b>There was no difference in polymorphism of IL-10-1082 between the endometriosis (AA: 87.90%, GA: 12.10%) and control group (AA: 87.50%, GA: 12.50%). The rate of TT, CT and CC genotype IL-10-819 was the same as the AA, CA and CC individually. There was no difference in the polymorphism of IL-10-819 or IL-10-592 between the endometriosis group (TT or AA: 41.12%, CT or CA: 47.66%, CC: 11.21%) and control group (χ(2) = 5.87, P = 0.053). However, there were significant difference in the genotype of CT of IL-10-819 or CA of IL-10-592 between the endometriosis group and control group (after adjust OR = 1.88, 95% CI = 1.10 - 3.21, χ(2) = 5.24, P = 0.021), and the allele C of IL-10-819 or IL-10-592 were close related with occurrence of endometriosis (OR = 1.42, 95%CI = 1.04 - 1.95, χ(2) = 4.81, P = 0.028). The IL-10 level in the plasma of endometriosis group with genotype of CC (CC), CT (CA) of IL-10-819(-592) were significant higher than those with TT (AA) (CA/CT: (50.12 ± 82.40) pg/ml, CC: (91.00 ± 118.23) pg/ml, TT/AA: (21.45 ± 22.10) pg/ml) (F = 2.492, P = 0.048; F = 1.852, P = 0.008).</p><p><b>CONCLUSION</b>The allele C of IL-10-819 or IL-10-592 was close related to the high level expression of IL-10, and it is the risk of the occurrence of endometriosis.</p>

Influence of SNCG transfection on the effectiveness of anti-tumor drugs in PC-3 cell lines / 中华男科学杂志

<p><b>OBJECTIVE</b>To observe the sensitivity of the PC-3 cell lines transfected with the PCI-NEO-SNCG plasmid to Cisplatin (DDP), 5-Fluorouracil (5-FU), Adriamycin (ADM), Vincristine (VCR) and Paclitaxel (TAX), and to explore the influence of the SNCG expression on the effectiveness of anti-tumor drugs.</p><p><b>METHODS</b>The plasmids PCI-NEO and PCI-NEO-SNCG were transfected into the hormone-independent prostate cancer cell lines PC-3. RT-PCR was adopted to examine the expression of SNCG in the PC-3 cell lines. The MTT method was employed to detect the suppressive effects of different anti-tumor drugs (DDP, ADM, 5-FU, VCR and TAX) on the cell lines transfected with PCI-NEO and PCI-NEO-SNCG. Flow cytometry was used to analyze the cell cycles and apoptosis of the transfected cells treated with TAX.</p><p><b>RESULTS</b>The 5 anti-tumor drugs suppressed the growth of the cell lines transfected with the plasmids PCI-NEO and PCI-NEO-SNCG in a time-dependant manner. The comparison between the growth-suppressing effects of different anti-tumor drugs on the PC-3 cell lines showed no significant differences between the group transfected with PCI-NEO and that with PCI-NEO-SNCG in DDP, 5-FU, ADM and VCR (P > 0.05), while the rate of suppression of TAX on the latter cell lines was significantly lower than that on the former (P < 0.01). Compared with the PCI-NEO-SNCG plasmid transfected cell lines, after treated with TAX for 48 hours, those transfected with the PCI-NEO plasmid exhibited a significantly larger proportion of cells remaining in the G2-M stage (P < 0.01), a smaller proportion in the G0-G1 and S stages (P < 0.01) and a significantly higher expression of Caspase-3 (P < 0.01).</p><p><b>CONCLUSION</b>The significant reduction of the growth-suppressing effect of TAX in the SNCG-transfected PC-3 cell lines suggests that the expression of SNCG may restrain the effect of TAX. These findings have provided evidence and guide to the individual chemotherapy of prostate cancer.</p>

Epidermal growth factor up-regulates the mRNA expression of endothelin-1 and its receptors in prostate cancer PC-3 cell lines / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the effects of the epidermal growth factor on the mRNA expression of endothelin-1 and its receptors (ET(A)R, ET(B)R) in hormone refractory prostate cancer (HRPC) PC-3 cell lines.</p><p><b>METHODS</b>PC-3 cells were cultured in vitro. After the treatment with EGF, the mRNA expressions of endothelin-1, ET(A)R and ET(B)R were detected by RT-PCR in PC-3 cell lines. The levels of the mRNA expression of endothelin-1 and its receptors were examined at different time points by RT-PCR.</p><p><b>RESULTS</b>The expressions of endothelin-1 and ET(A)R mRNA but not the mRNA expression of ET(B)R was observed in PC-3 cell lines. After 24 hours of treatment with EGF, the expressions of endothelin-1 and ET(A)R in PC-3 cell lines were both up-regulated and there was significant difference (P < 0.05) between the experimental and control groups. Different expression levels of endothelin-1 and ET(A)R mRNA were noted at different time points of EGF intervention, up-regulated with the increase of treatment time, and with significant difference (P < 0.05).</p><p><b>CONCLUSION</b>EGF can up-regulate the mRNA expressions of endothelin-1 and ET(A)R in PC-3 cell lines and play a great role in prostate cancer progression, which may offer a substructure of molecular biology for the treatment of HRPC.</p>

Advances in researches on the relationship between single nucleotide polymorphism and prostate cancer / 中华男科学杂志

Single nucleotide polymorphism (SNP) has been regarded as the third generation of heredity markers with many marked characteristics. It has been developed many new experimental techniques for detecting SNP in recent years, such as TaqMan probe technique, gene chip technique, denaturing high performance liquid chromatograph, matrix assisted laser desorption ionization-time off light mass spectrometry and minisequencing technique. Great progress has been made in researches on human tumors with SNP as heredity markers. This article reviews the genes and SNP related to the development of prostate cancer, including prostate specific antigen response component, enzymes, hormones and their receptors, cell cycle regulating protein, cytokines, adhesion molecules, vitamins and so on, and aims to explore the possible mechanism of the disease.

Expression of somatostatin receptor reporter gene and its correlation with targeted imaging in vivo for detection of pancreas carcinoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To evaluate the value of (99 m)Tc-sandostatin scintigraphy in targeted diagnosis of pancreas carcinoma and the correlation with expression of somatostatin receptor (SSTR) reporter gene in tumor tissue.</p><p><b>METHODS</b>Nude mice bearing human pancreas carcinoma xenograft were established in advance. (99 m)Tc-sandostatin imaging was performed in 18 nude mice and tumor to normal tissue ratio (T/NT) was calculated. mRNA expression of SSTR1, SSTR2 and SSTR5 in tumor tissue was detected by RT-PCR.</p><p><b>RESULTS</b>Out of 18 nude mice, 13 mice showed intense uptake of (99 m)Tc-sandostatin. Six hours after (99 m)Tc-sandostatin administration, the T/NT ratio was 2.53 +/- 0.84. Five mice showed negative findings, and the T/NT ratio was 1.04 +/- 0.06. Positive correlations were obtained between the T/NT ratio and the expression of SSTR1 and SSTR2 mRNA, especially SSTR2 (r = 0.807, P < 0.01).</p><p><b>CONCLUSION</b>High expression of SSTR1, SSTR2 and SSTR5 mRNA were found in human pancreas tumor xenograft in nude mice, especially SSTR2. There is a significant correlation between the tumor uptake of (99 m)Tc-sandostatin and SSTR2 mRNA level. This approach may allow SSTR-targeted diagnosis and therapy of human pancreas cancer.</p>

Detection and clinical significance of phospholipase A2 in semen of male infertile patients / 中华男科学杂志

<p><b>OBJECTIVES</b>To explore the clinical application of anti-human seminal plasma phospholipase A2 (PLA2) monoclonal antibody (McAb) for male infertility.</p><p><b>METHODS</b>Enzyme-linked immunoabsorbent assay (ELISA), immunocytochemistry(ICC), as well as flow cytometry (FCM) analysis were established using two strains anti-human seminal plasma PLA2 McAb prepared by our laboratory to detect the PLA2 content in human seminal plasma and the anterior head region of spermatozoa, respectively. Then the PLA2 content in male infertile patients were compared with that in normal control with fertility. The seminal routine analysis was performed by computer-assisted semen analysis (CASA).</p><p><b>RESULTS</b>The PLA2 content of infertile groups were (31.13 +/- 14.49) ng/ml in azoospermic patients, (17.71 +/- 12.45) ng/ml in oligospermic patients and (16.46 +/- 11.31) ng/ml in patients with normal sperm density, which were all higher than that of normal controls [(8.09 +/- 3.15) ng/ml, P < 0.01]; There was significantly negative correlation between PLA2 content in seminal plasma and sperm density(r = -0.602, P < 0.05), while there was insignificant correlation between PLA2 and sperm motility or percentage of motility. The PLA2 content in the anterior head region of spermatozoon of male infertile groups was significantly lower than that of normal controls by ICC and FCM(P < 0.01).</p><p><b>CONCLUSIONS</b>PLA2 in human seminal plasma is closely related to male fertility, and the PLA2 deficiency in the head of spermatozoa may be one of the reasons causing male infertility. The methods detecting PLA2 content in seminal plasma and the head of spermatozoa can provide powerful evidences for exploring the mechanism of male infertility.</p>

Detection of sexual hormone in semen of patients with idiopathic azoospermia or oligospermia and its significance / 中华男科学杂志

<p><b>OBJECTIVES</b>To detect the sexual hormone level in semen of patients with idiopathic azoospermia and oligospermia, and further analyze the relationship between sexual hormone and idiopathic azoospermia and oligospermia.</p><p><b>METHODS</b>50 male patients with idiopathic azoospermia, 50 in idiopathic oligospermia and 50 male controls with normal sperm density were selected. The sperm density and sexual hormone in semen were detected respectively by routine semen analysis and chemical luminescence technique.</p><p><b>RESULTS</b>The values of LH were (5.19 +/- 0.67) IU/L and (4.77 +/- 0.68) IU/L, and those of FSH were (1.90 +/- 0.79) IU/L and (2.27 +/- 0.25) IU/L in idiopathic azoospermia and oligospermia respectively, and the values of LH and FSH were (2.19 +/- 0.22) IU/L and (1.61 +/- 0.14) IU/L in normal control group respectively. There were significant differences in the values of LH and FSH between idiopathic azoospermia and normal control group(P < 0.01 or P < 0.05). The values of PRL were (6.25 +/- 0.51) ng/ml and (6.33 +/- 0.34) ng/ml, and those of T were (1.51 +/- 0.12) ng/ml and (1.68 +/- 0.71) ng/ml in idiopathic azoospermia and oligospermia respectively, and the values of PRL and T were (6.36 +/- 0.32) ng/ml and (1.83 +/- 0.09) ng/ml in normal control group respectively. There were no significant difference in the values of PRL between idiopathic azoospermia, oligospermia and normal control group, but there were significant differences of T between idiopathic azoospermia and normal control. Compared with 0.84 +/- 0.20 in normal control, the values of T/LH were 0.35 +/- 0.09 and 0.29 +/- 0.04 in idiopathic oligospermia and azoospermia respectively and there were significant differences(P < 0.05).</p><p><b>CONCLUSIONS</b>The changes of LH, FSH and T values may be one of the reasons that cause the dysfunction of spermatogenesis and sperm maturation in patients with idiopathic azoospermia and oligospermia. The study of semen hormone may lead to new strategies in the treatment to azoospermia and oligospermia.</p>

Construction and identification for cell strain of anti-human seminal plasma phospholipase A2 monoclonal antibody / 中华男科学杂志

<p><b>OBJECTIVES</b>To establish and evaluate the anti-human seminal plasma phospholipase A2 (PLA2) monoclonal antibody (McAb).</p><p><b>METHODS</b>After having been separated and purified from human seminal plasma by PEG precipitation, Sephacryl S-300 column chromatography, DEAE-Sephadex A-25 column chromatography and HA column chromatography, PLA2 was regarded as an antigen to immune BALB/C mouse to produce anti-human seminal plasma PLA2 McAb. The PLA2 McAb sensitivity and specificity were performed by ELISA technique and Western-blot analysis, respectively.</p><p><b>RESULTS</b>The molecular weight of PLA2 depurated with 245 fold purification from human seminal plasma was about 34,900, while the sensitivity and typing of its McAb were 1:5(6)-1:5(8) and IgM (kappa) with a satisfied Western-Blot results.</p><p><b>CONCLUSIONS</b>The PLA2, which had not been reported in international and domestic papers, may be a new type of PLA2. The establish of its McAb will provide significant tools for the research of the relationship between PLA2 in human seminal plasma and male fertility.</p>