Cell block technology in the pathological diagnosis of pleural effusion

Transcription

1 Chinese Journal of Tissue Engineering Research ( ) DOI: /j.issn ORCID: () (1) 105 (2) :R318 :B %(32/105) 44.8%(47/105) 57.1%(60/105) ( ) Cell block technology in the pathological diagnosis of pleural effusion Zhao Ye, Zhang Ji-xin, Liang Li, Li Ting (Department of Pathology, Peking University First Hospital, Beijing , China) Abstract BACKGROUND: Cytologic smear is a routine detection method for pleural effusion, but the results are far from satisfactory. Therefore, pleural effusion is made into cell block for immunochemistry and gene detection, to improve the detection accuracy, which contributes to the treatment and prognosis of pleural effusion. OBJECTIVE: To explore the application value of cell block technology in the pathological diagnosis and molecular detection of pleural effusion. METHODS: Totally 105 cases of pleural effusion samples were collected at Department of Pathology, Peking University First Hospital between January 2016 and August Effective components extracted after centrifugation were used to make paraffin blocks for hematoxylin-eosin staining and immunocytochemical staining thereafter. Genetic mutations were further detected in the cases of lung cancer diagnosed by cytology. Zhao Ye, Technician, Department of Pathology, Peking University First Hospital, Beijing , China Corresponding author: Li Ting, Master, Professor, Chief physician, Department of Pathology, Peking University First Hospital, Beijing , China 1934 : (2018)

2 Zhao Y, Zhang JX, Liang L, Li T. Cell block technology in the pathological diagnosis of pleural effusion. Zhongguo Zuzhi Gongcheng Yanjiu. 2018;22(12): DOI: /j.issn RESULTS AND CONCLUSION: The positive rate of routine cytological smear was 30.5% (32/105), which was significantly lower than that of the cell block section (44.8%, 47/105). The positive rate of immunocytochemical staining combined with cell block was the highest one with 57.1%, 60/105). Besides, this method could be used to identify tumor types. Gene mutation detection was performed in 19 cases of lung cancer, among which 13 cases were positive. These results indicate that cell block technology combined with immunocytochemical staining is an effective method for the diagnosis of malignancy and tumor origin in pleural effusion, and can be further used for mutation detection. Subject headings: Pleural Effusion; Cytological Techniques; Immunohistochemistry; Pathology, Molecular; Neoplasms Funding: the National Natural Science Foundation of China, No Introduction 48.6% [1] 17% [2] [3] [4] [5] 1Materials and methods ml h50 ml2 000 r/min 5 min(5% 15%) 1015 ml( )10 min 95%10 min r/min10 min 310% 1 h 4 µm (1) µm 60 1 h (Autostainer Link-Thermofish)( Dako Denmark A/S) EnVision [6] EGFRK-rasALK ROS1 8 µm 10 EGFR/K-rasFFPE DNACatNO.ADx-FF01ALK/ROS1 FFPE RNACatNO.ADx-FF04 P.O. Box 10002, Shenyang

3 . [J] (12): DOI: /j.issn A B C A B 1 Figure 1 Procedures of making cell blocks AC 2 ( 400) Figure 2 Immunocytochemistrical staining results of lung adenocarcinoma cells ( 400) AB NapsinA TTF1 A B [n(%)] Table 1 Comparison of the diagnosis results of the cytologic smear, cell block and the combination of cell block and immunocytochemical staining 3 ( 400) Figure 3 Immunocytochemistrical staining results of the mantle cell lymphoma ( 400) AB CD20 CyclinD1 63(60.0) 58(55.2) a 45(42.9) b 32(30.5) 47(44.8) a 60(57.1) b 10(9.5) a P < Table 2 Characteristics of the cells diagnosed with the combination of cell block and immunocytochemical staining [n(%)] P < (42.9) 27(25.7) 3(2.9) 3(2.9) 1(1.0) 1(1.0) 1(1.0) 1(1.0) 3(2.9) 2(1.9) 18(17.1) Nanodrop 2000DNA OD260 nm/od280 nm RNA 260 nm/280 nm ARMS PCREGFR(ADx-EG01) K-ras(ADx-KR05-MX)ALK(ADx-AE02)ROS1 (ADx- RO02)stratagene Mx3000P PCR( )2 PCR SPSS 2 Results (60.0%)32(30.5%)10(9.5%) (44.8%) TCT(P < 0.05) 105TCT 45(42.9%)60(57.1%)(1) (P < 0.05) 27(2)3 ()3 11(3) (2) ISSN CN /R CODEN: ZLKHAH

4 Zhao Y, Zhang JX, Liang L, Li T. Cell block technology in the pathological diagnosis of pleural effusion. Zhongguo Zuzhi Gongcheng Yanjiu. 2018;22(12): DOI: /j.issn %53.3% 60.0% EGFRKRASALKROS EGFR[ (L858R)]2 EML4-ALKK-rasROS1 3Discussion [7] 23 TTF-1Napsin A [8] CD20B [9] CyclinD1 [10] [11] TCT32(30.5%) (57.1%60/105)(100%) 76.9%100% 50%100% [7] 105 [12] 19 EGFRALK ROS1K-ras1911EGFR2 EML4-ALKK-rasROS1 EGFR-T790M37.5% [13] [14-24] ( 18 h) ( ) (STROBE ) CNKI 3 () ISSN CN /R CODEN: ZLKHAH 1937

5 . [J] (12): DOI: /j.issn References [1],,,.EGFR19 PCR[J].,2006,26(7): [2],,,.142 [J]., 2017,33(3): 292. [3],,,. [J].,2016,32(12): [4],,,. [J]., 2014, 22(12): [5],. [J].,2017,22(6): [6],.[J]., 2017,22(11): [7] Ugurluoglu C, Kurtipek E, Unlu Y, et al. Importance of the cell block technique in diagnosing patients with non-small cell carcinoma accompanied by pleural effusion. Asian Pacific journal of cancer prevention: APJCP. 2015;16(7): [8] Turner BM, Cagle PT, Sainz IM, et al. Napsin a, a new marker for lung adenocarcinoma, is complementary and more sensitive and specific than thyroid transcription factor 1 in the differential diagnosis of primary pulmonary carcinoma: evaluation of 1674 cases by tissue microarray. Arch Pathol Lab Med. 2012;136(2): [9].[J]., 2010,17(1):4-6. [10] Werling RW, Yaziji H, Bacchi CE, et al. CDX2, a highly sensitive and specific marker of adenocarcinomas of intestinal origin: an immunohistochemical survey of 476 primary and metastatic carcinomas. Am J Surg Pathol. 2003;27(3): [11] Fetsch PA, Abati A. Immunocytochemistry in effusion cytology: a contemporary review. Cancer. 2001;93(5): [12] Cheng F, Wang Q, Zhong D. Value of Cell Block in the Diagnosis of Malignant Pleural Effusion. Zhongguo Feiai Zazhi. 2015;18(10): [13] Satouchi M, Tanaka H, Yoshioka H. Detection of epidermal growth factor receptor gene T790M mutation in cytology samples using the cobas EGFR mutation test. Lung Cancer. 2017;111: [14] Wang W, Tang Y, Li J, et al. Detection of ALK rearrangements in malignant pleural effusion cell blocks from patients with advanced non-smallcell lung cancer: a comparison of Ventana immunohistochemistry and fluorescencein situ hybridization. Cancer Cytopathol. 2015;123(2): [15] Billon P, Bryant EE, Joseph SA, et al. CRISPR-Mediated Base Editing Enables Efficient Disruption of Eukaryotic Genes through Induction of STOP Codons. Mol Cell. 2017;67(6): [16] Paolillo C, Mu Z, Rossi G, et al. Detection of Activating Estrogen Receptor Gene (ESR1) Mutations in Single Circulating Tumor Cells.Clin Cancer Res. 2017;23(20): [17] Haque F, Varlet P, Puntonet J, et al. Evaluation of a novel antibody to define histone 3.3 G34R mutant brain tumours. Acta Neuropathol Commun. 2017;5(1):45. [18] Willems H, Jacobs A, Hadiwikarta WW, et al. Thermodynamic framework to assess low abundance DNA mutation detection by hybridization. PLoS One. 2017;12(5):e [19] Wang W, Yuan Y, Zheng H, et al. A Pilot Study of Noninvasive Prenatal Diagnosis of Alpha- and Beta-Thalassemia with Target Capture Sequencing of Cell-Free Fetal DNA in Maternal Blood. Genet Test Mol Biomarkers. 2017;21(7): [20] Janku F, Zhang S, Waters J, et al. Development and Validation of an Ultradeep Next-Generation Sequencing Assay for Testing of Plasma Cell-Free DNA from Patients with Advanced Cancer. Clin Cancer Res. 2017;23(18): [21] Fricke F, Lee J, Michalak M, et al. TGFBR2-dependent alterations of exosomal cargo and functions in DNA mismatch repair-deficient HCT116 colorectal cancer cells. Cell Commun Signal. 2017;15(1):14. [22] Spira A, Yurgelun MB, Alexandrov L, et al. Precancer Atlas to Drive Precision Prevention Trials. Cancer Res. 2017;77(7): [23] Kasahara N, Kenmotsu H, Serizawa M, et al. Plasma epidermal growth factor receptor mutation testing with a chip-based digital PCR system in patients with advanced non-small cell lung cancer. Lung Cancer. 2017;106: [24] Breveglieri G, Travan A, D'Aversa E, et al. Postnatal and non-invasive prenatal detection of β-thalassemia mutations based on Taqman genotyping assays. PLoS One. 2017;12(2): e ISSN CN /R CODEN: ZLKHAH