Features of genes copy number in normal, primary tumor, and lymph nodes metastases cells in patients with cervical cancer.

e17503 Background: Early stages of cervical cancer (CC) can be treated with surgery or radiation therapy, metastatic CC is incurable, and therefore new diagnostic approaches based on highly effective molecular markers are required. As such markers, the Copy Number Variation (CNV) is of great interest. Purpose of the study: to analyze the peculiarities of genes CNV in normal cells, primary tumor and lymph nodes tumor metastases cells in patients with CC. Methods: Tissue sections from FFPE-blocks of 300 patients with CC were used for the study. Primary tumor, tumor metastatic and normal cells were isolated using non-contact laser microdissection (Palm MicroBeam, Carl Zeiss). DNA was extracted from the cells using the phenol-chloroform method. Determination of 14 genes CNV (LAMP3, PRKAA1, TORC2, FOXO3, HDAC5, MEF2C, MLXIPL, EP300, HNF4A, TP53, SREBF1, SREBF2, PPARGC1A, and CCND1) was performed by Real-Time qPCR method. Statistical analysis was performed using the Mann-Whitney test; Bonferroni's correction was applied to correct multiple comparisons. Results: An increase in the CNV of LAMP3 and TORC2 genes by 2.5 and 3.2 times (p < 0.05), respectively, and a decrease in the CNV of TP53 and FOXO3 genes by 2.3 and 2.0 times (p < 0.05), respectively, were found in the primary tumor cells relative to normal cells. The effect of the simultaneous change in the CNV of LAMP3, TORC2, TP53 and FOXO3 genes was observed in 60% of the sample, and LAMP3 and TP53 genes - in 80%. In cells of lymph nodes tumor metastases an increase in the CNV of CCND1 and TORC2 genes by 2.0 and 3.5 times (p < 0.05), respectively, was found, as well as a decrease in the CNV of PPARGC1A gene by 2.1 times (p < 0.05) relative to normal cells. A simultaneous change in the CNV of CCND1, TORC2, and PPARGC1A was observed in 70% of the sample, and CNV of CCND1 and TORC2 genes - in 85%. Primary tumor and metastases cells differed in the CNV of CCND1 (2.5 times higher in metastatic cells), LAMP3 (3.0 times lower in metastatic cells) and FOXO3 genes (2.0 times higher in metastatic cells). Conclusions: Thus, the primary tumor and lymph nodes metastases cells differ in the level of genes CNV from normal cervix cells. Accordingly, the CNV of LAMP3, TORC2, TP53 and FOXO3 genes may have the potential for diagnosing a primary cervical tumor, and the CNV of CCND1, TORC2 and PPARGC1A genes for diagnosing its metastases to regional lymph nodes.

Value of MRI and diffusion weighted imaging in diagnosing normal-sized pelvic lymph nodes metastases in patients with cervical cancer

Objectives: To investigate the value of conventional magnetic resonance imaging (MRI) and diffusion weighted imaging (DWI) in diagnosing normal-sized pelvic lymph nodes metastases in patients with cervical cancer. Methods: 102 patients with cervical cancer who underwent MRI and DWI scan were included. 137 lymph nodes were analyzed, including 44 metastatic lymph nodes(MLNs) and 93 non-metastatic lymph nodes(non-MLNs).The morphology and apparent diffusion coefficient (ADC) value of lymph nodes were measured including short-axis diameter (DS), long-axis diameter (DL), ratio of short to long-axis diameter (DR), fatty hilum, asymmetry, ADCmax, ADCmean and ADCmin. The Mann-Whitney U test, independent sample t test and Chi-square test were employed to compare the difference of all criteria between MLNs and non-MLNs. Logistic regression and decision tree were used to develop the combined diagnostic model. ROC analyses were used to evaluate the diagnostic performance. Results: The DS and DR of MLNs were significantly higher than those of non-MLNs (p < 0.05), the ADCmax, ADCmean and ADCmin of MLNs were significantly lower than those of non-MLNs (p < 0.05). Presence of fatty hilum and asymmetric lymph nodes between MLNs and non-MLNs were significantly different (p＜0.05). Combined measurement of ADCmin, DS and DR had the highest AUC 0.937 with 90.9% sensitivity and 87.1% specificity. The accuracy of decision tree was 88.3%. Conclusion: MRI with DWI had potential in diagnosing normal-sized pelvic lymph nodes metastases in patients with cervical cancer. The combined evaluation of DS, DR and ADCmin of lymph nodes and decision tree of the combined measure showed better diagnostic performances than sole criteria. Advances in knowledge: The short-axis diameter, ratio of short to long-axis diameter and ADCmin of lymph nodes have moderate value in the diagnosis of the metastases of the normal-sized lymph nodes for the patient with cervical cancer as the sole indices. The combined evaluation of DS, DR and ADCmin is much more valuable in the detection of metastatic lymph nodes.