HOXC6 and HOXC8 are potentially novel prognosis predictors of esophageal squamous cell carcinoma
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1 HOXC6 and HOXC8 are potentially novel prognosis predictors of esophageal squamous cell carcinoma Keneng Chen, M.D., PhD, RCSF Luyan Shen, M.D., PhD 2015/4/28 Department of Thoracic Surgery I, Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education), Beijing Cancer Hospital, Peking University School of Oncology, Beijing, China
2 No potential conflicts of interest were disclosed
3 Several issues with the clinical practice of ESCC remain unsolved 01 The morbidity and mortality remains high 02 long-term survival remains unsatisfied 03 Lack of reports with long-term follow-up data from prospective database 04 Lack of efficient biomarkers for helping to guide the clinical practice prognosis
4 Why we choose HOX genes? 01 There is some similarity between embryogenesis and tumorigenesis 02 AFP in liver cancer and CEA in colorectal cancer 03 HOX gene family regulates embryogenesis 04 HOX genes are deregulated in hematologic malignancy and many solid tu
5 Regional expression of transcription factors in the developing gut Yuasa Y. Nat Rev Cancer
6 Chen KN, et al. Clin Cancer Res Expression of 11 HOX genes is deregulated in ESCC HOX genes were abnormally expressed in ESCC (n=36) Gene Positive cases in ESCC(%) HOXA10 3(8.3) 0(0) HOXA13 27(75) 0(0) HOXB7 21(58.3) 0(0) HOXC4 12(33.3) 0(0) HOXC8 15(41.7) 0(0) HOXD9 6(16.7) 0(0) HOXD10 15(41.7) 0(0) HOXD13 15(41.7) 0(0) HOXA7 30(83.3) 12(33.3) HOXA9 24(66.7) 9(25) HOXC6 24(66.7) 9(25) Positive cases in noncancerous mucosa(%) potential biomakers
7 Study design and work flow diagram FFPE tissue specimen Lentivirus infection KYSE-450 IHC for HOXC6 HOXC8 protein detection S Group (138 ) NC+S Group (136) S+AC Group (148) CCK8 Assay Annexin v/pi Soft agar colony formation assay Prognosis Proliferation Cell cycle & Apoptosis Ability of colony formation Knockdown of HOXC6/HOXC8 Tumor formation in nude mice Ability of tumorigenesis
8 Staining of IHC
9 Survival analysis in surgery subgroup Surgery only p = p = The association of HOXC6/HOXC8 expression with overall survival in naive subgroup (n=138) Item No. (%) Median survival, 5-y survival p mo(95%ci) (%) HOXC6 Low expression 83(60.1) 70.1( ) High expression 55(39.9) 35.0( ) 32.9 HOXC8 Low expression 85(61.6) 110.3( ) High expression 53(38.4) 30.3( ) 30.6 Du YB, et al. J Surg Res, 2014.
10 Survival analysis in neoadjuvant chemotherapy subgroup p = p = NC+S The association of HOXC6/HOXC8 expression with overall survival in neoadjuvant subgroup (n=136) Item No.(%) Median survival mo(95%ci) 5-y survival (%) HOXC6 Low expression 31(22.8) 82.7(-) High expression 105(77.2) 27.8( ) 33 HOXC8 Low expression 73(53.7) 110.3(-) High expression 63(46.3) 21.2( ) 25 Du YB, et al. J Surg Res, p
11 Survival analysis in adjuvant chemotherapy subgroup p = p = S+AC The association of HOXC6/HOXC8 expression with overall survival in neoadjuvant subgroup (n=148) Item No.(%) Median survival 5-y survival p mo(95%ci) (%) HOXC6 Low expression 81(54.7) 61.0( ) High expression 67(45.3) 33.4( ) 30.3 HOXC8 Low expression 99(66.9) 62.3( ) High expression 49(33.1) 31.3( ) 21.7
12 HOXC6/HOXC8 Knock-down at mrna and Protein level mrna Protein
13 Down-regulation inhibited cell proliferation in vitro & in vivo CCK8 assay shnc Tumor growth in nude mice shhoxc6 shhoxc8 Soft agar Colony formation HOXC6(-) HOXC8(-) Control
14 Down-regulation induced cell apoptosis Apoptosis rate variation when HOXC6 or HOXC8 was knockdown Apoptosis rate(%) shhoxc6 32.8±0.29 * shhoxc8 36.1±0.35 * ShNC 12.7±0.31 * p < 0.050
15 Down-regulation arrested cell cycle in G1 phase HOXC6(-) HOXC8(-) Control Knockdown of HOXC6 and HOXC8 Affects the Cell Cycle G2+S(%) G1(%) shhoxc ±2.29 * 54.62±2.29 * shhoxc ±3.1 * 71.9±3.1 * shnc 64.87± ±3.17 * p < 0.050
16 Conclusion 01 HOXC6, HOXC8 might be biomarker for evaluating prognosis of ESCC 02 HOXC6/HOXC8 promotes the esophageal cancer cell proliferation
17 Acknowledgment Dr. Chen s LAB: Yabing Du Bin Dong Wanpu Yan Liang Dai Xiaozheng Kang Zhen Liang Hongchao Xiong
18
19 HOX genes present the expression pattern of temporal and spatial colinearity A13 A11 A10 A9 B9 A6 A5 A4 A3 A2 A1 7p15.3 B13 B9 B8 B7 B6 B5 B4 B3 B2 B1 17p21.3 C13 C12 C11 C10 C9 C8 C6 C5 C4 12q13.3 D13 D12 D11 D10 D9 D8 D4 D3 D1 2q31 Posterior Late ' lumbo-sacral thoracic cervical 3' Anterior Early Schematics representation of four HOX loci Cillo C, et al. J cell physiol, 2001
20 HOXC6/HOXC8 expression was related with TNM stage Association between HOXC6 or HOXC8 expression and clinical characteristics (n=274) Clinicopathologic Characteristics HOXC6 expression No.(%) P HOXC8 expression No.(%) P High Low High Low Age 60 74(62.7) 44(37.3) (44.9) 65(55.1) >60 87(55.0) 71(45.0) 63(39.9) 95(60.1) Sex Male 121(59.0) 84(41.0) (42.9) 117(57.1) Female 40(56.3) 31(43.7) 28(39.4) 43(60.6) G stage G1 10(41.7) 14(58.3) (33.3) 16(66.7) G2 25(39.0) 39(61.0) 29(45.3) 35(54.7) G3 20(40.0) 30(60.0) 16(32.0) 34(68.0) Tumor location(l) L0 15(62.5) 9(37.5) (37.5) 15(62.5) L1 38(67.9) 18(32.1) 26(46.4) 30(53.6) L2 75(59.1) 52(40.9) 57(44.9) 70(55.1) L3 30(46.2) 35(53.8) 23(35.4) 42(64.6) Tumor invasion(t) T1 33(47.1) 37(52.9) (27.1) 51(72.9) <0.001 T2 33(50.0) 33(50.0) 22(34.9) 41(65.1) T3 68(63.0) 40(37.0) 52(48.1) 56(51.9) T4 26(78.8) 7(21.2) 23(69.7) 10(30.3) Lymph node metastasis (N) N0 96(53.9) 82(46.1) (36.5) 113(63.5) N1 33(58.9) 23(41.1) 32(57.1) 24(42.9) N2 20(71.4) 8(28.6) 10(35.7) 18(64.3) N3 11(91.7) 1(8.30) 9(75.0) 3(25.0) TNMG stage I 79(45.7) 94(54.3) < (34.1) 114(65.9) II 41(73.2) 15(26.8) 28(50.0) 28(50.0) III 41(87.2) 6(12.8) 29(61.7) 18(38.3)
21 Tumor Regression Grade (TRG) NC+S The association of HOXC6 and HOXC8 expression with TRG (n=136) HOXC6 expression No.(%) p HOXC8 expression No.(%) p Low High Low High TRG TRG1/2 15(48.4) 27(30.9) (47.9) 7(11.1) <0.001 TRG3/4 16(51.6) 78(69.1) 38(52.1) 56(88.9) Du YB, et al. J Surg Res, 2014.
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