scholarly journals Predictive Value of Circulating miRNAs in Lymph Node Metastasis for Colon Cancer

Genes ◽  
2021 ◽  
Vol 12 (2) ◽  
pp. 176
Author(s):  
In Hee Lee ◽  
Gyeonghwa Kim ◽  
Sang Gyu Kwak ◽  
Dong Won Baek ◽  
Byung Woog Kang ◽  
...  
Keyword(s):  
Colon Cancer ◽  
Lymph Node ◽  
Cancer Patients ◽  
Circulating Mirnas ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Diagnosis And Prognosis ◽  
Polymerase Chain ◽  
Validation Set ◽  
Plasma Mirna

(1) Background: Lymph node (LN) status is an indubitable prognostic factor for survival among colon cancer patients. MicroRNAs (miRNAs) have been implicated in the development and progression of many cancers and are potential biomarkers for cancer diagnosis and prognosis. Therefore, we validated candidate biomarkers using circulating miRNAs by analyzing the plasma miRNA concentrations from patients with colon cancer to predict LN metastasis. (2) Methods: This study included 79 blood samples from patients diagnosed with colon cancer. The NanoString assay was used for screening, and TaqMan miRNA assays for quantitative real-time polymerase chain reaction (RT-PCR) test was used for validation. In a discovery set, we compared the expression of 800 circulating miRNAs in 24 samples (stage 0/I/IIA versus IIIB/IIIC). For validation, a total 79 samples were tested using quantitative RT-PCR. (3) Results: In the discovery set, 10 candidate circulating miRNAs were detected (4 up-regulated miRNAs: miR-323a-3p, miR-382-5p, miR-29a-3p, and miR-376a-3p; 6 down-regulated miRNAs: miR-26a-5p, let-7g-5p, miR-15b-5p, miR-142-3p, miR-374a-5p, and let-7b-5p). In the validation set, higher expression of three circulating miRNAs (miR-323a-3p, miR-382-5p, and miR-376a-3p) was significantly associated with LN metastasis (p = 0.0063, 0.0107, and 0.0022). (4) Conclusions: High expression of circulating miR-323a-3p, miR-382-5p, and miR-376a-3p was significantly associated with LN metastasis in colon cancer patients. These miRNAs could be circulating biomarker candidates that predict the presence of LN metastasis.

Immunohistochemistry and Reverse Transcriptase Polymerase Chain Reaction on Sentinel Lymph Nodes can Improve the Accuracy of Nodal Staging in Breast Cancer Patients

2001 ◽  
Vol 16 (4) ◽  
pp. 227-232 ◽  
Author(s):  
G. Péley ◽  
J. Tóth ◽  
O. Csuka ◽  
I. Sinkovics ◽  
E. Farkas ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Polymerase Chain Reaction ◽  
Lymph Node ◽  
Lymph Nodes ◽  
Reverse Transcriptase ◽  
Sentinel Lymph Nodes ◽  
Nodal Staging ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain

In this study the nodal staging sensitivity of sentinel lymph node biopsy (SLNB) with detailed pathological and molecular biological examination has been investigated and compared to that of axillary lymph node dissection (ALND) with routine histological evaluation. Sentinel lymph nodes (SLNs) were removed by the dual-agent injection technique in 68 patients with primary, clinically node-negative breast cancer. Forty-seven patients had negative SLNs according to hematoxylin and eosin (H&E) staining. These H&E-negative SLNs were serially sectioned and examined at 250 μm levels by anticytokeratin immunohistochemistry (IHC). In 14 patients the SLNs were also investigated by cytokeratin 20 (CK20) reverse transcriptase polymerase chain reaction (RT-PCR). SLNB with IHC increased the node-positive rate by 26% (by 40% in tumors less than or equal to 2 cm in size (pT1) and by 9% in tumors more than 2 cm but less than or equal to 5 cm in size (pT2)). The sensitivity of SLNB with IHC was superior to that of ALND with routine histology in pT1 tumors and identical in pT2 tumors. The concordance between histology and RT-PCR was only 21%, and in two of three cases with positive histological results RT-PCR was negative. In conclusion, SLNB with detailed pathological and/or molecular biological evaluation can improve the sensitivity of regional staging. ALND can probably be abandoned in patients with pT1 SLN-negative breast cancer. Further prospective studies are required to determine the clinical significance of these detailed SLN evaluation techniques, but at present these methods are still investigational.

Sentinel lymph node micrometastasis detection using immunohistochemistry and reverse transcription-polymerase chain reaction for cytokeratin 19 in breast cancer

2007 ◽  
Vol 25 (18_suppl) ◽  
pp. 11074-11074
Author(s):  
Q. Cao ◽  
S. Wang ◽  
X. Liu ◽  
L. Lin ◽  
J. Zhao
Keyword(s):  
Breast Cancer ◽  
Polymerase Chain Reaction ◽  
Lymph Node ◽  
Sentinel Lymph Node ◽  
Detection Rate ◽  
Cytokeratin 19 ◽  
Breast Cancer Patients ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain

11074 Background: In breast cancer, the sentinel lymph node (SLN) can be used to assess the axillary nodal status to guide the axillary surgery, tumor staging and adjuvant systematic therapy. Histology and immunohistochemistry (IHC) are currently the routine methods of SLN assay. The facts that about 30% of node-negative breast cancer patients relapse within five years and that micrometastasis were found in 9%∼30% negative lymph nodes when they were re-examined using serial sectioning suggest that current histological detection methods are inadequate for identifying metastatic tumor cells in lymph nodes. The primary objective of this study was to develop an assay by using a combination of reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry for cytokeratin 19 (KT19) expression in SLN. Methods:139 SLN samples were collected from 139 breast cancer patients undergoing SLN biopsy using Isosulfan Blue dye during modified mastectomy between June 2002 and June 2005. The SLNs were dissected into two parts, half were frozen in liquid nitrogen during sugery for RT-PCR detection and another half were kept in liquor formaldehyde for routine hematoxylin-eosin (H&E) examination and IHC detection. In 62 tumor-negative SLNs at routine H&E examination (pN0), we performed IHC and RT-PCR for KT19. Results: Of the 62 tumor-negative SLNs at routine H&E examination, 11% (7/62) were positive on extensive IHC analyses for KT-19, which were all positively identified by RT-PCR assay. Six SLNs (10%) were negative for micrometastases (with H&E and IHC for KT-19) but RT-PCR positive. There was asignificant difference in detective rate between these two methods statistically (χ2 = 4.1667 , P = 0.0412) and the coincidence were high ( kappa = 0.6483) Conclusions: As IHC analysis resulted in a 11% detection rate in H&E negative SLNs, IHC is essential to avoid false- negative SLN micrometastases. Molecular analysis with KT19 allows detection of breast cancer micrometastases with an additional 10% detection rate in H&E negative SLNs and resulted in a high coincidence with IHC analysis. However, the clinical value of these histologically negative but RT-PCR positive SLNs can only be determined with long term follow up. No significant financial relationships to disclose.

scholarly journals Polyisoprenoids from Avicennia marina induces on P13k, Akt1, Mammalian target of rapamycin, Egfr, and P53 Gene Expression Using Reverse Transcription-Polymerase Chain Reaction

2020 ◽  
Vol 8 (A) ◽  
pp. 146-152
Author(s):  
Taufiq Qurrohman ◽  
Mohammad Basyuni ◽  
Poppy Anjelisa Zaitun Hasibuan
Keyword(s):  
Gene Expression ◽  
Colon Cancer ◽  
Polymerase Chain Reaction ◽  
Reverse Transcription ◽  
Mammalian Target Of Rapamycin ◽  
P53 Gene ◽  
Avicennia Marina ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain

BACKGROUND: According to the Global Cancer Observatory in 2018, Asia was the first to note the incidence of colon cancer, which was 51.8% of cases of colon cancer which ranked the top three in the number of causes of death in the world. Cancer is a disease characterized by uncontrolled cell growth. Potential natural ingredient developed as chemotherapeutic agents includes from mangrove leaves. Studies reporting on the pharmacological activity of polyisoprenoid from mangrove species are still limited, therefore, it is essential to achieve the prospects, potential, and mechanisms polyisoprenoid in mangroves as a natural ingredient of pharmaceutical and medication. AIM: The aim of the study was to investigate the inhibition activities of polyisoprenoids in mangrove plant Avicennia marina in WiDr cells induces on P13k, Akt1, mammalian target of rapamycin (mTOR), Egfr, and P53 gene expression using reverse transcription-polymerase chain reaction (RT-PCR). MATERIAL AND METHODS: The leaves of A. marina were dried and extracted with n-hexane followed by evaporation and freeze-drying. Polyisoprenoid contents were analyzed with two-dimensional thin-layer chromatography method. Cell viability was assessed with 3-(4,5-dimetiltiazol-2-il)-2,5-diphenyl tetrazolium bromide assay. The cycle cell was tested with flow cytometry method. The apoptotic test was determined with a double-staining method. The gene expression on P13k, Akt1, mTOR, Egfr, and P53 was analyzed by RT-PCR method. RESULTS: The results showed that 48 h cytotoxic activity of polyisoprenoids against WiDr cells and 5-Fu (positive control) had IC50 values, 295.25 μg/mL and 17.43 μg/mL. Cell cycle analysis depicted that the inhibition of polyisoprenoid occurred in the G0-G1 phase and 5-Fu in S phase. Polyisoprenoid and 5-Fu had the same mechanism in the early apoptotic phase. RT-PCR revealed that polyisoprenoids downregulated the P13k, Akt1, mTOR, and Egfr gene expression, and however, upregulated P53 gene expression. CONCLUSION: The present study confirmed that polyisoprenoids from A. marina leaves showing as chemopreventive agents for colon cancer.

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