Clinical applicability of xTAG liquidchip technology as EGFR mutations detection method in lung cancer / 临床与实验病理学杂志

Purpose To analyze EGFR exon 18 ~21 gene mutations in lung cancer, and compare xTAG liquidchip technology and Sanger sequencing technology in clinical practice. Methods 1 139 tumor tissue samples from phaseⅠtoⅣlung cancer patients were randomly collected. DNA was extracted from the samples. EGFR gene mutation status in exon 18~21 was detected by xTAG liquidchip technology and Sanger sequencing technology respectively. Results The mutation status of EGFR was obtained by xTAG liquidchip technology in 1 134 patients, and 1 105 by Sanger sequencing technology, detection success rate was 99. 56% and 97. 01% respective-ly. The sensitivity and specificity of xTAG liquidchip technology Comparing with Sanger sequencing was 99. 59% and 94. 54%. Sever-al cases of multiple mutations were detected by both methods. All mutation types detected by two methods are fully consistent. Conclu-sions Comparing with Sanger sequencing technology, xTAG liquidchip technology, which is able to detect mutations of exon 18~21 simultaneously, is more convenient and efficient for EGFR gene mutation detection in lung cancer.

Serum miR-103 as a potential diagnostic biomarker for breast cancer / 南方医科大学学报

<p><b>OBJECTIVE</b>To evaluate the diagnostic value of serum miR-103 for breast cancer and its correlation to the clinicopathological features of the patients.</p><p><b>METHODS</b>We collected the serum samples and corresponding formalin-fixed paraffin-embedded (FFPE) surgical specimens from 50 breast cancer patients, using the serum samples from 50 healthy women as the control. The total RNA was extracted from the samples for quantitative analysis of miR-103 using real-time RT-PCR.</p><p><b>RESULTS</b>The serum levels of miR-103 expression were significantly higher in the cancer patients than in the healthy control group (P<0.01). In the cancer patients, high miR-103 expression was significantly correlated to advanced clinical stage (P<0.05) and lymph node metastasis (P<0.05). miR-103 showed a receiver operating characteristic (ROC) curve area of 84.3%, and a sensitivity of 84% and a specificity of 70% in discriminating breast cancer patients from the control subjects.</p><p><b>CONCLUSION</b>Serum miR-103 can serve as a potential diagnostic marker for breast cancer and provides useful information of the clinicopathological features of the patients.</p>

High frequency loss of heterozygosity on the long arms of chromosomes 13 and 14 in nasopharyngeal carcinoma in Southern China / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To investigate the loss of heterozygosity (LOH) on chromosomal arms 13q and 14q in nasopharyngeal carcinoma (NPC) using 21 microsatellite polymorphic markers and to study whether there is a correlation between LOH and clinicopathologic parameters and/or Epstein-Barr virus (EBV) infection in NPC.</p><p><b>METHODS</b>Sixty cases of NPC were studied using polymerase chain reaction based microsatellite analysis with genescan and genotyping techniques.</p><p><b>RESULTS</b>LOH was detected on 13q in 78% of NPC tumors, high frequency LOH loci (more than 30%) clustered to 13q12.3-q14.3 and 13q32. On chromosome 14q, LOH was detected in 80% of NPC tumors; high frequency LOH loci clustered to 14q11-q13, 14q21-q24 and 14q32. High frequency LOH at 13q31-q32 correlated with a lower level of EBV infection; LOH on chromosome 14q was closely associated with poor differentiation of NPC tumor cells.</p><p><b>CONCLUSION</b>Our results suggest that in NPC, LOH on chromosome 13q and 14q are common genetic events, and putative tumor suppressor genes (TSG) residing in these regions may be involved in tumorigenesis.</p>

The construction of lentivirus-mediated RNAi vector containing cytochrome C oxidase / 中国病理生理杂志

AIM: To construct a recombinant lentivirus RNAi vector carrying cytochrome C oxidase gene to obtain the titer of the lentiviral stock for investigation of the expression in the eukaryotic cell and the affection of the COX gene silencing in the eukaryotic cells. METHODS: According to the DNA of the cytochrome C oxidase gene, we designed and synthesized complementary single-strand DNA oligos, annealed the single-stranded oligos to generate a ds oligo, cloned the ds oligo into pENTR/U6 to obtain an entry clone; An LR recombination reaction was performed between the pENTR/U6 entry construct and pLenti6/BLOCK-iT-Dest to generate expression construct, the 293FT cell line was cotransfected with pLenti6/BLOCK-iT expression construct, and the viral packaging mix, viral supernatant was harvested to determine the titer. RESULTS: The DNA sequence of interest clone to the vector was constructd to generate an entry clone and an expression clone successfully, which were proved by sequence determination. A vector producing cell line 293FT was established, and the titer for transfection was obtained. Western blotting analysis demonstrated that COX shRNA expression construction could suppress the expression of MTCOX-I. CONCLUSION: A lentivirus RNAi vector containing cytochrome C oxidase gene was successfully constructed.