Cancer Biomarkers

Thus far, biological roles of plasminogen activator inhibitor 1 (PAI1) in hepatocellular carcinoma (HCC) remain controversial. Moreover, its expression, clinicopathologic and prognostic significance in HCC have not been comprehensively investigated, therefore needing further evidence.

PAI1 expression was measured, using tissue microarray-based immunohistochemical staining, in matched HCC and adjacent liver samples from 178 patients with HCC after curative resection. The correlations of PAI1 H-scores with clinicopathologic variables and survival were further evaluated. Its prognostic value was finally confirmed in some public databases.

It was found that PAI1 expression was significantly higher in HCC than in adjacent liver tissues. Moreover, high PAI1 expression was more frequent in those with multiple lesions. Univariate analyses showed that PAI1 expression was negatively associated with both overall and relapse-free survival. Although PAI1 expression was not statistically significant in multivariate Cox regression test, combination of it with TNM stage effectively distinguished survival and relapse, and served as an independent prognostic factor. In the online available datasets of HCC and liver cancer used, SERPINE1, the gene encoding PAI1, was also revealed to be prognostic.

Our data suggested that high PAI1 expression was predictive for unfavorable biological behavior and long-term prognosis in HCC.

Anastomotic leak (AL), as one of the most devastating complications, is the leading cause of mortality in colorectal cancer (CRC) patients after resection. This study was aimed to investigate potential risk factors for AL in elderly surgical CRC patients.

A total of 1068 elderly subjects who underwent elective curative colorectal surgery from 2012 to 2018 were retrospectively evaluated and enrolled into this study population. The predictive value of C-reactive protein-to-albumin ratio (CAR) for AL in surgical CRC patients was evaluated by receiver operating characteristic (ROC) curve analysis. Potential risk factors for AL were assessed by the univariate and multivariate logistic regression analyses.

Of all the 1068 enrolled patients, 81 patients have developed AL with an incidence of 7.6% (81/1068). Preoperative CAR was an effective predictor for AL with an area under the curve (AUC) of 0.758, 95% CI of 0.700–0.817, a cut-off value of 2.44, a sensitivity of 61.09% and a specificity of 80.25%, respectively ( P < 0.001). Duration of operation (OR: 2.05, 95% CI: 1.21–3.44, P = 0.013) and preoperative CAR (OR: 1.94, 95% CI: 1.21–3.11, P = 0.007) were two independent risk factors for AL by the multivariate logistic regression analysis.

Our study indicate that preoperative CAR level and duration of operation were two independent predictors for AL among elderly surgical CRC patients.

The current cervical cancer screening strategies based on Papanicolaou (Pap) and Human papillomavirus (HPV) tests receive great achievement but still exhibit many limitations in clinical practice. Exploring new biomarkers as stratified management method in HPV primary screening is becoming the tendency of current research.

Immunocytochemistry (ICC) of FHIT and C-MYC were performed on exfoliated cervical cells from 197 eligible high-risk HPV positive women. Mann-Whitney U test, Pearson Chi-Square test, logistic regression analysis and receiver operating characteristic (ROC) curves were used to assess the diagnostic efficiency.

ICC staining intensity of FHIT and C-MYC in high-grade cervical intraepithelial neoplasia (CIN) specimens was significantly different from low-grade CIN and normal specimens. Compared with Pap test, ROC analysis of ICC in detecting high-grade CIN resulted in a larger area under the curve (AUC) (0.805 and 0.814 vs 0.723, p < 0.001). FHIT achieved higher sensitivity than Pap test (79.41% vs 66.67%, p = 0.04). Logistic regression analysis of the combination of two biomarkers led to higher AUC value, specificity and PPV than any single biomarker.

The utility of FHIT and C-MYC ICC analysis in cervical exfoliated cells of HPV-positive women displayed superior diagnostic potential and may improve clinical performance of cervical cancer screening.

Chemotherapy remains the basis of the treatment of lung cancer, and screening biomarkers with predictive value for chemotherapy is of great interest. The present study focused on status of genes methylation in NSCLC patients receiving pemetrexed- or gemcitabine-based chemotherapy.

Promoter methylation of Ras association domain family (RASSF1A) and short stature homeobox 2 (SHOX2) was examined in bronchoalveolar lavage (BAL) from 117 NSCLC patients treated with chemotherapy. Multivariate analysis was used to identify the predictive value of gene methylation. Progression-free survival (PFS) rather than overall survival (OS) was used as the clinical outcome to minimize the impact of chemotherapy on gene methylation.

The methylation of RASSF1A and SHOX2 was significantly associated with shorter PFS (RASSF1A: HR = 2.355, 95% CI: 1.533–3.617, P < 0.0001; SHOX2: HR = 2.123, 95% CI: 1.392–3.236, P = 0.0004). After adjusting for confounding factors, RASSF1A methylation was still a predictive factor for PFS (HR = 1.765, 95% CI: 1.064–2.928, P = 0.0278). In the pemetrexed group, unmethylated RASSF1A could be used to predict longer PFS ( P = 0.0001), and no predictive value was found in the gemcitabine group.

Unmethylated RASSF1A is a favorable prognostic indicator for patients receiving pemetrexed doublets. Because of the promoting effect of most chemotherapeutic drugs on gene methylation, unmethylated RASSF1A is not suitable as a predictor for gemcitabine doublets.

To study the relationship between the tumor heterogeneity based on CT and overall survival (OS) in oesophageal squamous cell carcinoma treated with chemotherapy and radiation therapy (CRT).

Fifty-seventh clinical patients who underwent definitive CRT were analyzed. The results were analyzed in terms of whole-tumor texture, with quantification of entropy, mean gray-level intensity for fine to coarse textures (filters 1.0–2.5, respectively). The association between texture parameters and survival time was assessed by Kaplan-Meier analysis and a Cox proportional hazards model.

The median, 1 and 3 years OS, were 20.2 months, 75.4%, and 32.1%. In the univariate analysis performed using the log-rank test found global entropies ( P = 0.0119), global mean ( P = 0.088), global Std ( P = 0.0209), and global uniformity ( P = 0.0284) were found to be significant OS prognostic factors for filter value 1.0. Cox proportional hazards models that included a combination of pretreatment GlobalStd and post-treatment volume yields the best performance in predicting OS. Kaplan-Meier curves show that the patients in the high-risk group have significantly worse OS (log-rank test, P = 0.0009) and progression-free survival (PFS) (log-rank test, P = 0.0019) than those in the low-risk group.

Pretreatment texture parameters are associated with survival time, and the combination of post volume performed better in survival models.

The key role in carcinogenesis with destruction of the extracellular matrix is played by proteases released by invasive cancer cells. Cysteine peptidases, such as cathepsin B and L, take an important role in cancer progression and metastasis.

Cysteine peptidase-like activity (CPA) in sera of patients with breast cancer at different stages of disease and the influence of genetic predisposition associated with BRCA-1 gene mutations were analysed.

CPA in serum was determined with the spectrofluorometric technique using Z-Phe-Arg-AMC as a substrate. Determination was carried out in 111 breast cancer patients in comparison to a control group of 50 healthy subjects.

The highest CPA was found in breast cancer patients with a hereditary predisposition bearing BRCA1 gene mutations, and the lowest activity was found in patients who had a tumour surgically removed and before adjuvant therapy. The differences in the activities between control group and cancer groups were statistically significant ( p < 0.05), except from group of cancer patients in complete remission ( p < 0.52).

Serum CPA in patients with breast cancer differs depending on the cancer stage and treatment methods. Our study demonstrate the correlation between BRCA-1 gene mutations and the increased level of CPA.

Uveal melanoma (UM) is the most common primary intraocular tumor in adults, which has a high mortality rate and worse prognosis. Therefore, early potential molecular detection and prognostic evaluation seem more important for early diagnosis and treatment.

Gene expression data were obtained from The Cancer Genome Atlas-Uveal melanomas database. Survival genes were identified by univariate analysis and were regarded to be associated with the overall survival of UM patients. Then, pathway enrichment analysis of these survival genes was performed. Robust likelihood-based survival model and multivariate survival analysis were conducted to identify more reliable genes and the prognostic signature for UM survival prediction. Two internal datasets and another two UM datasets from Gene Expression Omnibus (GEO) were used for the validation of prognostic signature.

Firstly, 2,010 survival genes were screened by univariate survival analysis. GO and KEGG analysis revealed that these genes were mainly involved in pathways such as mRNA processing, RNA splicing, spliceosome and ubiquitin mediated proteolysis. Secondly, a six-gene signature was identified by Robust likelihood-based survival model approach. The gene expression of the six genes can successfully divide UM samples into high- and low-risk groups and have strong survival prediction ability. What’s more, the expression of six genes was compared in 80 healthy adipose tissue samples obtained from GTEx (Genotype-Tissue Expression) database and further validated in internal datasets and GEO datasets, which also can predict UM patient survival.

The six genes (SH2D3A, TMEM201, LZTS1, CREG1, NIPA1 and HIST1H4E) model might play a vital role in prognosis of UM, which should be helpful for further insight into the treatment of uveal melanoma.

Previous studies have showed that long non-coding RNAs (lncRNAs) are critical regulators in many cancers. The aim of this study is to investigate the clinical role and functional effects of long non-coding RNA SNHG14 in pancreatic ductal adenocarcinoma (PDAC).

The expression of SNHG14 in 58 pairs of pancreatic cancer tissues and adjacent normal tissues was detected by quantitative real-time PCR (qRT-PCR) analysis. The correlations between SNHG14 expression and PDAC patients’ clinicopathological characteristics and prognosis were statistically assessed. Cell counting kit-8 (CCK8) and transwell cell invasion assays were employed to detect the capacities of cell proliferation and cell invasion. The western blot analysis was used to detected the expression of E-cadherin and Vimentin.

In the study, we found that SNHG14 expression was higher in PDAC tissue compared to adjacent normal tissues by qRT-PCR analysis. Higher SNHG14 expression was significantly associated with advanced TNM stage and positive lymph node metastasis in PDAC patients. Furthermore, we demonstrated that higher SNHG14 expression acted as a poor predictor in PDAC patients compared with lower SNHG14 expression. Moreover, we showed that higher SNHG14 expression promoted cell proliferation, cell colony formation and cell invasion ability in PDAC. Upregulation of SNHG14 expression promoted cell invasion by affecting E-cadherin expression via interacting with EZH2.

Thus, these results indicated that SNHG14 expression acts as a prognostic maker for PDAC and potential target of PDAC treatment.

METCAM/MUC18 expression was increased with the malignant progression of prostate cancer and also a bona fide metastatic gene, capable of initiating and driving the metastasis of a non-metastatic human prostate cancer cell line to multiple organs.

We explored if METCAM/MUC18 was detectable in human serum and a novel biomarker to predict malignant propensity of prostate cancer.

Two antibodies were identified by Western blot analysis having the highest sensitivity and specificity to establish calibration curves from the recombinant METCAM/MUC18 proteins. They were used in ELISA and LFIA to determine the METCAM/MUC18 concentrations in serum samples from 8 normal individuals, 4 BPH patients, 1 with PIN, 6 with high-grade prostate cancer, and 2 treated cancer patients.

Serum METCAM/MUC18 concentrations were statistically significantly higher in the patients with PIN and prostate cancer than those with BPH, the treated patients and normal individuals. The LFIA results were statistically better than ELISA and Western blot methods. Serum METCAM/MUC18 concentrations were in direct proportional to most of serum PSA concentrations.

Pancreatic cancer is one of the most lethal malignancies worldwide. Pancreatic adenosquamous carcinoma (PASC) is a rare histological type of pancreatic carcinoma with a poor prognosis. The median survival time after diagnosis is less than one year. It is believed that the pathogenesis of PASC is different from pancreatic adenocarcinoma. In this study, we tried to reveal the intrinsic gene mutations associated with PASC through whole exome sequencing.

Both cancerous and paracancerous tissues were collected from 12 pathologically diagnosed PASC patients. Their clinical characteristics were collected, and patient survival information was obtained through follow-up. The correlations between the mutations and clinical characteristics were analysed.

Germline mutations were identified in MAP3K1 (9 cases), PDE4DIP (7), BCR (7), ALK (6), USP6 (5), AR (4), HLA-A (4), SPEN (4), KMT2D (3), NUTM2B (3), ZFHX3 (3), and MN1 (3), while somatic mutations were found in TP53 (5), KRAS (3), HRNR (3), and OBSCN (3). Peripheral tissue invasion was associated with somatic mutations in KRAS ( P = 0.0339). Additionally, there were significant correlations between lymphatic metastasis and germline mutations in USP6 ( P = 0.0228) and somatic mutations in OBSCN and HRNR ( P = 0.0339).

In conclusion, susceptibility genes including MAP3K1, PDE4DIP, and BCR are frequently found to be mutated in the germlines of PASC patients. Somatic mutations in KRAS, OBSCN, and HRNR and germline mutations in USP6 are related to tumour invasion and metastasis, reinforcing the necessity of translating these potential biomarkers into clinical practice.

The majority of ovarian cancer cases are diagnosed at an advanced stage with poor prognosis. This study evaluates autoantibodies against tumor antigens to identify candidate biomarkers for early detection of ovarian cancer in women at increased risk.

To assess the immunoreactivity of paraneoplastic antigens and tumor associated antigens with high-grade serous ovarian cancer (HGSOC) samples.

Five paraneoplastic antigens along with three tumor-associated antigens were evaluated with HGSOC patient serum samples. Validation screening was performed with n = 164 serum samples consisting of: 50 late stage HGSOC, 14 early stage HGSOC, 50 benign ovarian cyst, and 50 healthy control samples on ELISA and western blot. The four markers TRIM21, NY-ESO-1, TP53, and PAX8 were evaluated on a second validation serum set, n = 150.

TRIM21 achieved the highest sensitivity in the first validation screening of 33% with 100% specificity. Combining TRIM21 with NY-ESO-1, TP53, and PAX8 provided 67% sensitivity with 94% specificity, and 56% sensitivity at 98% specificity. These four markers resulted in 46% sensitivity with 98% specificity in the second validation cohort; TRIM21 achieved the highest individual sensitivity of 36%.

Autoantibodies to TRIM21, NY-ESO-1, and TP53 may complement CA125 in screening of women at genetic risk for ovarian cancer.