The Combining Effects of Cell-Free Circulating Tumor DNA of Breast Tumor to the Noninvasive Prenatal Testing Results: A Simulating Investigation.

Massively parallel sequencing of circulating fetal DNA in the plasma of pregnant women is a common method for noninvasive prenatal testing (NIPT) of fetal trisomy 13, 18, and 21. However, circulating DNA is not restricted to pregnant women, with increased levels of plasma DNA also frequently detected in the plasma of cancer patients. Among pregnant women whose NIPT results were inconsistent with the fetal karyotype, a small number of patients have subsequently been diagnosed with a previously undetected malignancy. However, the extent to which circulating tumor DNA (ctDNA) affects the results of NIPT is still unclear. We examined serum from 50 nonpregnant women with breast tumors by NIPT. These samples were then added to serum containing trisomy 13, 18, and 21 fetal DNA to figure out the extent to which maternal tumors can interrupt NIPT results in pregnant women with breast tumors. Concentrations of cell-free DNA (cfDNA) were higher in both pregnant women and breast tumor patients, relative to nonpregnant healthy controls. Among the 50 samples evaluated, 3 produced false positive NIPT results for trisomy 13, 18, or 21, indicating that genomic copy number variations (CNVs) had occurred. Simulation testing also showed that ctDNA can increase the standard deviation of the associated z-scores, which lower absolute z-scores by decreasing the proportion of circulating fetal DNA relative to total DNA. Of the 50 samples tested, 9 fell within the equivocal range and 8 produced false negative results for trisomy 13, 18, or 21. Data presented here show for the first time that ctDNA is able to affect NIPT results in two ways. First, ctDNA can lead to false positive results due to the detection of genomic CNVs in tumor DNA. Alternatively, ctDNA can increase the likelihood of a false negative by decreasing the proportion of circulating fetal DNA in serum.

Endoscopic naso-leakage drainage: a safe and effective method for the management of intrathoracic anastomotic leakage after esophagectomy.

BACKGROUND: Intrathoracic anastomotic leakage (IAL) remains a major complication of esophagectomy. Main non-surgical options of management include chest drainage and endoscope interventions. This study is aim to present our experience and assess the efficacy of endoscopic naso-leakage drainage (ENLD) in patients with IAL. METHODS: From June 2011 to January 2017, 67 patients who developed IAL after esophagectomy and managed by non-surgical approaches were analyzed retrospectively. IAL was confirmed by clinical presentations combined with the evidence of CT scan, radiography and endoscopy. Thirty-eight patients were treated by conventional chest drainage (CD group) and 29 patients underwent ENLD with or without chest drainage (ENLD group), while other treatments including enteral nutrition and antibiotics had no difference between the two groups. In ENLD group, a 12 Fr naso-leakage tube was placed through the leakage to the bottom of vomica under ultra-slim electronic gastroscope. The naso-leakage tube was then connected to a gastrointestinal decompression device for drainage and was also used for rinse. When the vomica diminished and the drainage was also clean, the naso-leakage tube could be pulled back gradually. Finally, healing of the leakage was confirmed endoscopically. Clinical records of the two groups were analyzed. RESULTS: In ENLD group, naso-leakage tubes were successfully placed under endoscope in all 29 patients without any procedure-related complications. In CD group, the mortality is 7.9% (three patients) and five patients (13.2%) developed to systemic inflammatory response syndrome (SIRS) due to insufficient drainage. While in ENLD group, there was only one patient (3.4%) developed to SIRS and no death was observed, but the difference was not statistically significant. When compared with the CD group, the ENLD group had a shorter healing course (44.2±18.3 vs. 60.5±27.7 days, P=0.008), duration of antibiotics usage (16.4±7.8 vs. 11.8±3.8 days, P<0.001) and duration of fever (4.3±2.2 vs. 9.5±8.6 days, P=0.002). CONCLUSIONS: To our initial experience, ENLD is an ideal option with safety and efficacy in management of IAL after esophagectomy.

Neoadjuvant Bevacizumab plus Chemotherapy versus Chemotherapy Alone to Treat Non-Metastatic Breast Cancer: A Meta-Analysis of Randomised Controlled Trials.

PURPOSE: Results from previous randomised controlled trials (RCTs) investigating whether the addition of bevacizumab to neoadjuvant chemotherapy (NAC) could statistically significantly increase the pathological complete response (pCR) and to identify which subgroup would benefit most from such regimens have produced conflicting results. This meta-analysis was designed to assess the efficacy and safety of bevacizumab plus chemotherapy compared with chemotherapy alone in the neoadjuvant setting. METHODS: A literature search of MEDLINE, EMBASE, Web of Science, and the Cochrane library was performed to identify eligible studies. The primary endpoint of interest was pCR. The secondary endpoints were clinical complete rate (cCR), surgery rate, breast-conserving surgery (BCS) rate, and toxicity. The meta-analysis was performed using Review Manager software version 5.3. RESULTS: Nine RCTs matched the selection criteria, yielding a total of 4967 patients (bevacizumab plus chemotherapy: 50.1%, chemotherapy alone: 49.9%). The results of this meta-analysis demonstrated that the addition of bevacizumab to NAC significantly increased the pCR rate (odds ratio [OR] = 1.34 [1.18-1.54]; P < 0.0001) compared with chemotherapy alone. Subgroup analysis showed that the effect of bevacizumab was more pronounced in patients with HER2-negative cancer (OR = 1.34 [1.17-1.54]; P < 0.0001) compared with HER2-positive cancer (OR = 1.69 [0.90-3.20]; P = 0.11). Similarly, in patients with HER2-negative cancer, the effect of bevacizumab was also more pronounced in patients with HR-negative cancer (OR = 1.38 [1.09-1.74]; P = 0.007) compared with HR-positive cancer (OR = 1.36 [0.78-2.35]; P = 0.27). No significant differences were observed between the groups with respect to cCR, surgery rate, or BCS rate. Additionally bevacizumab was associated with a higher incidence of neutropenia, febrile neutropenia, and hand-foot syndrome. CONCLUSIONS: Higher proportions of patients achieved pCR when bevacizumab was added to NAC compared with when they received chemotherapy alone; acceptable toxicities were also found. Subgroup analysis demonstrated that patients with histologically confirmed HER2-negative and HR-negative breast cancer benefited the most.

CHRNA3 polymorphism modifies lung adenocarcinoma risk in the Chinese Han population.

Recent genome-wide association studies (GWASs) have identified 15q25.1 as a lung cancer susceptibility locus. Here, we sought to explore the direct carcinogenic effects of genetic variants in this region on the risk of developing lung adenocarcinoma (ADC). Five common SNPs (rs8034191, rs16969968, rs1051730, rs938682, and rs8042374) spanning the 15q25.1 locus were assayed in a case-control study examining a cohort of 301 lung ADCs and 318 healthy controls. Stratification analysis by gender, smoking status, and tumor, node, metastasis (TNM) classification, was performed. In addition, sections from ADC tissue and normal tissue adjacent to tumors were stained with an anti-CHRNA3 (cholinergic receptor nicotinic α3) antibody by immunohistochemistry in 81 cases. Our results demonstrate that rs8042374, a variant of the CHRNA3 gene, is associated with an increased risk of ADC with an OR of 1.76 (95% CI: 1.17-2.65, p=0.024). This variant was linked to a greater risk of ADC in female nonsmokers (OR (95% CI): 1.81 (1.05-3.12), p=0.032) and female stage I+II cases (OR (95% CI): 1.92 (1.03-3.57), p=0.039). Although located within the same gene, rs938682 showed protective effects for smokers, stage III+IV cases, and male stage III+IV cases. Additionally, the CHRNA3 protein level in ADC tissue was slightly higher than in the surrounding normal lung tissue, based on immunohistochemical analysis. Our results suggest that the CHRNA3 polymorphism functions as a genetic modifier of the risk of developing lung ADC in the Chinese population, particularly in nonsmoking females.

Breast cancer association studies in a Han Chinese population using 10 European-ancestry-associated breast cancer susceptibility SNPs.

BACKGROUND: Genome-wide association studies (GWAS) have identified various genetic susceptibility loci for breast cancer based mainly on European-ancestry populations. Differing linkage disequilibrium patterns exist between European and Asian populations. METHODS: Ten SNPs (rs2075555 in COL1A1, rs12652447 in FBXL17, rs10941679 in 5p12/MRPS30, rs11878583 in ZNF577, rs7166081 in SMAD3, rs16917302 in ZNF365, rs311499 in 20q13.3, rs1045485 in CASP8, rs12964873 in CDH1 and rs8170 in 19p13.1) were here genotyped in 1009 Chinese females (487 patients with breast cancer and 522 control subjects) using the Sequenom MassARRAY iPLEX platform. Association analysis based on unconditional logistic regression was carried out to determine the odds ratio (OR) and 95% confidence interval (95% CI) for each SNP. Stratification analyses were carried out based on the estrogen receptor (ER) and progesterone receptor (PR) status. RESULTS: Among the 10 SNPs, rs10941679 showed significant association with breast cancer when differences between the case and control groups in this Han Chinese population were compared (30.09% GG, 45.4% GA and 23.7% AA; P = 0.012). Four SNPs (rs311499, rs1045485, rs12964873 and rs8170) showed no polymorphisms in our study. The remaining five SNPs showed no association with breast cancer in the present population. Immunohistochemical tests showed that rs2075555 was associated with ER status; the AA genotype showed greater association with ER negative than ER positive (OR = 0.54, 95% CI, 0.29-0.99; P = 0.046). AA of rs7166081 was also associated with ER status, but showed a greater association with ER positive than negative (OR = 1.59, 95% CI = 1.04-2.44; P = 0.031). However, no significant associations were found among the SNPs and PR status. CONCLUSION: In this study using a Han Chinese population, rs10941679 was the only SNP associated with breast cancer risk, indicating a difference between European and Chinese populations in susceptibility loci. Therefore, confirmation studies are necessary before utilization of these loci in Chinese.

Mutational analysis of key EGFR pathway genes in Chinese breast cancer patients.

BACKGROUND: The epidermal growth factor receptor (EGFR) is a potential therapeutic target for breast cancer; however, its use does not lead to a marked clinical response. Studies of non-small cell lung cancer and colorectal cancer showed that mutations of genes in the PIK3CA/AKT and RAS/RAF/MEK pathways, two major signalling cascades downstream of EGFR, might predict resistance to EGFR-targeted agents. Therefore, we examined the frequencies of mutations in these key EGFR pathway genes in Chinese breast cancer patients. METHODS: We used a high-throughput mass-spectrometric based cancer gene mutation profiling platform to detect 22 mutations of the PIK3CA, AKT1, BRAF, EGFR, HRAS, and KRAS genes in 120 Chinese women with breast cancer. RESULTS: Thirteen mutations were detected in 12 (10%) of the samples, all of which were invasive ductal carcinomas (two stage I, six stage II, three stage III, and one stage IV). These included one mutation (0.83%) in the EGFR gene (rs121913445-rs121913432), three (2.5%) in the KRAS gene (rs121913530, rs112445441), and nine (7.5%) in the PIK3CA gene (rs121913273, rs104886003, and rs121913279). No mutations were found in the AKT1, BRAF, and HRAS genes. Six (27.3%) of the 22 genotyping assays caused mutations in at least one sample and three (50%) of the six assays queried were found to be mutated more than once. CONCLUSIONS: Mutations in the EGFR pathway occurred in a small fraction of Chinese breast cancers. However, therapeutics targeting these potential predictive markers should be investigated in depth, especially in Oriental populations.

[Value of cardiac troponin I measurement in prediction of anthracycline-induced cardiotoxicity in breast cancer patients].

OBJECTIVE: To evaluate the value of cardiac troponin I (CTnI) measurement in predicting anthracycline-induced cardiotoxicity in patients with breast cancer. METHODS: This study was conducted among 186 breast cancer patients receiving anthracycline-based chemotherapy. Serum cTnI concentrations before and after each cycle of the chemotherapy and the left ventricular ejection fraction (LVEF) before and at the 2nd, 4th and 6th months of the treatment were recorded. According to serum cTnI concentration, the patients were divided into CTnI+ group (with serum CTnI concentration of no less than 0.1 ng/ml, n=60) and CTnI- (<0.1 ng/ml) group (n=126). RESULTS: No patients in this series experienced cardiac heart failure (CHF). The number of patients with a LVEF reduction by over 10% from the baseline was 16 (26.7%) in CTnI+ group, as compared to 7 (5.6%) in CTnI- group, showing a significant difference between the two groups (P<0.01). CONCLUSION: CTnI can be a useful marker for early prediction of anthracycline-induced cardiotoxicity in patients with breast cancer.

HLA class II variants in Chinese breast cancer patients.

Alterations of human leukocyte antigen (HLA) class II molecules are relevant to the development of breast cancer and metastatic progression. However, the role of HLA class II polymorphisms in the pathogenesis and progression of breast cancer is unclear. This study aimed to investigate the association between HLA class II variants and breast cancer susceptibility and prognosis in a Chinese population. Sixteen variants in HLA class II were detected with the Sequenom MassArray® iPLEX System in 216 breast cancer patients and 216 healthy controls. An association analysis based on unconditional logistic regression was carried out to determine the odds ratio (OR) and 95% confidence interval (95% CI) for each SNP. Stratified analysis by oestrogen receptor (ER) and progesterone receptor (PR) status was also performed. Among 16 variants, only seven conformed to Hardy-Weinberg proportions in the controls. None of these seven variants showed statistically significant differences between the case and control groups in this Han Chinese population. However, chr6_32737733, a variant in HLA-DQB1, showed significant associations with both ER-negative and PR-negative breast cancer in the best fit to the dominant model. Furthermore, another significant correlation was seen between chr6_32606112, a variant in HLA-DRB5, and PR positivity. These results indicate that although no breast cancer risk variants in HLA class II were found in this Chinese population, HLA-DQB1 chr6_32737733 may be involved in determining a poor prognosis, whereas HLA-DRB5 chr6_32606112 may relate to a good prognosis.

[Effect of cellular immune supportive treatment on immunity of esophageal carcinoma patients after modern two-field lymph node dissection].

BACKGROUND & OBJECTIVE: Cellular immunity suppression is marked in patients with esophageal carcinoma, which may be resulted temporarily from surgical injury. This study was to evaluate the effect of cellular immune supportive treatment on cellular immunity of patients with esophageal carcinoma. METHODS: A total of 60 patients with thoracic esophageal carcinoma, received two-field dissection, were randomized into control group and trial (immune supportive treatment) group. The patients in trial group were injected with Shenqi injection after operation; the patients in control group received no immune supportive treatment. Peripheral blood samples were obtained before operation, and 3 and 9 days after operation. AgNOR (argyrophilic nucleolar organizer regions) activity in peripheral blood T lymphocytes was measured by tumor immune microphotometry. T cell subsets were measured by flow cytometry. RESULTS: The proportions of CD3+CD4+ and CD4+/CD8+ cells were significantly higher in trial group than in control group at 3 days after operation (P < 0.05). The amount of AgNOR and proportions of CD3+, CD3+CD4+, CD4+/CD8+, and CD4+CD25+ cells were significantly higher in trial group than in control group at 9 days after operation (P < 0.05). There was no significant difference in 1-year survival rate between the 2 groups (P > 0.05). CONCLUSION: Shenqi injection could obviously improve cellular immunity of the esophageal carcinoma patients after modern two-field dissection.

[Effects of MT1-MMP on the in vitro invasiveness of breast cancer cells].

OBJECTIVE: To investigate the effect of membrane type-1 matrix metalloproteinase (MTI-MMP) on the invasive potential of breast cancer cell and analyze its mechanisms. METHODS: After treatment of breast cancer MDA-MB-453 cell line with concanavalin A ( ConA, 20 microg/ml) for 24 h, MT1-MMP protein was detected in cancer cells by Western analysis and immunocytochemistry. MDA-MB-453 cells were cultured with exogenous latent proMMP-2 and MMP-2 activity was analyzed by gelatin zymography. The invasive potential of the tumor cells was measured with a membrane invasion culture system. Cancer cells of the cell line were divided into four groups: the control group treated by neither reagent, group ConA was only treated by ConA, group MMP-2 was treated only by MMP-2, and group ConA + MMP-2 was treated by both ConA and MMP-2. RESULTS The expression of MTI-MMP protein could be detected in groups ConA and ConA + MMP-2, but nothing was detected in control and group MMP-2. There was only 72 000 precursor form of MMP-2 in group MMP-2 and there were both 72 000 precursor form and 64 000 active enzyme form of MMP-2 in group ConA + MMP-2, but there was no forms of MMP-2 in the other two groups detected by gelatin zymography. The largest amount of cells penetrated through Matrigel was observed in group ConA + MMP-2 than in the other three groups. CONCLUSION: MTI-MMP can remarkably promote the invasive potential of breast cancer cells mainly through its ability of activating latent proMMP-2 to degrade

[Significance of membrane type-1 matrix metalloproteinase expression in breast cancer].

BACKGROUND & OBJECTIVE: Recent researches manifested that MT1- MMP (Membrane type-1 matrix metalloproteinase) is essential for breast cancer invasion and metastasis, but there are few reports about the clinical value of MT1- MMP. This study was designed to investigate the clinical significance of MT1-MMP protein and mRNA expression in human breast cancers. METHODS: Forty-six human breast cancer tissues were collected. MT1-MMP protein and mRNA expression were detected by semi-quantitative reverse transcriptase chain reaction (RT-PCR) and immunohistochemistry assay and their correlations with clinicopathological factors were analyzed. RESULTS: The expression of MT1-MMP protein was positive in 52.2% of forty-six breast cancer tissues. The positive rates were 12.5%, 54.8% and 85.7% in stageI, stage II and stage III human breast cancers, respectively (P < 0.05), and 11.1%, 59.4% and 80.0% in the group of T(1), T(2) and T(3) (P < 0.05), and 27.3%, 66.7% and 100.0% in the group of N(0), N(1) and N(2) (P < 0.05). MT1-MMP mRNA expression were detected in all of the carcinomas. The means of MT1-MMP mRNA expression relative quantities were 0.4730, 0.6950 and 0.9100 in stageI, stage II and stage III human breast cancers, respectively (P < 0.05), and 0.5240, 0.7150 and 0.8220 in the group of T(1),T(2) and T(3) (P < 0.05), and 0.6305, 0.7022 and 0.8700 in the group of N(0), N(1) and N(2)(P < 0.05). There were no relations between the MT1-MMP protein or mRNA expression and ER, PR or C-erbB-2(P >0.05). CONCLUSION: MT1-MMP protein and mRNA expression in human breast cancers had positive correlation with the stage,the tumor size and lymph node metastasis. MT1-MMP can be a predictor for the ability of breast cancer invasion and metastasis.

[Expression correlation of Ki67 to P53,VEGF,and C-erbB-2 genes in breast cancer and their clinical significances].

BACKGROUND & OBJECTIVE: The proliferating antigen Ki67 is an important biological marker in cell proliferation. This study was designed to investigate the expressions of Ki67, P53, vascular endothelial growth factor (VEGF),and C-erbB-2 in breast cancer tissue, and their correlations with cliniopathological significance. METHODS: The expressions of Ki67,p53,VEGF,and C-erbB-2 in 151 cases of breast cancer were assessed by immunohisto-chemistry, and their correlations with clinicopathological factors were statistically analyzed. RESULTS: The positive rate of Ki67 was 78.2%(113/151). The expression of Ki67 correlated with tumor stages (P< 0.05), but didn't relate to age, tumor size, and lymph node status (P >0.05). In addition,the expression of Ki76 had a positive correlation with the expressions of p53, and C-erbB-2 (P< 0.05), but not with the expression of VEGF (P >0.05). The co-expression of Ki67 and VEGF related to tumor size,and clinical stage (P< 0.05). CONCLUSIONS: The expression of Ki67 is an objective biological marker for estimating the occurrence and progression of breast cancer. Combined detection of Ki67 and VEGF may helps to judge the clinical stage of breast cancer.

[Matrix metalloproteinases and breast cancer].

Matrix metalloproteinases(MMPs) are a family of zinc-dependent proteinases involved in the degradation of the extracellular matrix. A great deal of studies showed that they have the important role in invasiveness and metastasis of breast carcinoma. It was recently reported that they also contribute to the genesis and development of primary breast carcinoma. MMPs take part in these processes on various mechanisms. Clinically, MMPs showed values for treatment and judging prognosis of breast carcinoma. At present, the scholars are investigating the mechanisms of MMPs on breast carcinoma by various methods, so as to approach a new pathway for treating breast carcinoma.