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Journal of Chinese Integrative Medicine ›› 2012, Vol. 10 ›› Issue (4): 406-415.doi: 10.3736/jcim20120409

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Characteristic genomics of peripheral blood mononuclear cells of hepatocellular carcinoma patients with liver-kidney yin deficiency syndrome

Li Weng, Juan Du, Wen-ting He, Chang-quan Ling()   

  1. Department of Traditional Chinese Medicine, Changhai Hospital, Second Military Medical University, Shanghai 200433, China
  • Received:2011-10-25 Accepted:2011-12-23 Online:2012-04-20 Published:2018-04-15

Objective: To explore the characteristic genomics of syndrome of liver-kidney yin deficiency in peripheral blood mononuclear cells of hepatocellular carcinoma (HCC) patients.
Methods: HCC patients with or without syndrome of liver-kidney yin deficiency were enrolled into the experimental group and the control group, respectively; their gene expression profiles were evaluated by a whole-genome Affymetrix GeneChip Human Genome U133 Plus 2.0 Array. The differentially expressed mRNAs were then selected by Gene Ontology (GO) and pathway analyses, respectively. Based on the results of GO and pathway analyses, gene coexpression networks were built according to the normalized signal intensity of specifically expressed genes. Finally, the results from microarray were confirmed by real-time fluorescence quantitative polymerase chain reaction and Western blot methods.
Results: The results showed that a set of 615 mRNAs were differentially expressed in the HCC patients with syndrome of liver-kidney yin deficiency. By GO enrichment analysis, the genes for anti-apoptosis, regulation of cell cycle, transmembrane transport, etc. were up-regulated or down-regulated in the experimental group. Another functional analysis of mRNAs by KEGG revealed that 10 signal transduction pathways were up-regulated and 16 were down-regulated, such as antigen processing and presentation, cell cycle, and protein export. Based on the above results, we constructed coexpression networks to determine which genes may play pivotal role in HCC patients with syndrome of liver-kidney yin deficiency. Some critical genes, including SEC62 (SEC62 homolog (S. cerevisiae)), CCNB1 (cyclin B1) and BIRC3 (baculoviral IAP repeat-containing 3), which rank the top 3 in |Δ normalized degree| were chosen. Of another 60 samples, we found that the mRNA expressions of SEC62, CCNB1 and BIRC3were significantly lower in HCC patients with syndrome of liver-kidney yin deficiency than those without syndrome of liver-kidney yin deficiency (P<0.01). Also, the protein expressions of SEC62, CCNB1 and BIRC3 were significantly lower (P<0.01).
Conclusion: Gene chip technique allows rapid and high-throughput screening for different gene expression in HCC patients with or without liver-kidney yin deficiency syndrome. The results of this study further confirm the hypothesis on the essence of syndrome, namely, a kind of deviation from the normal state in multigene style on the levels of both mRNA and protein.

Key words: liver neoplasms, liver-kidney yin deficiency, gene expression, genomics

Table 1

Primer sequences and length of products"

Gene Upstream Downstream Length (bp)
SEC62 5′-AGCTGACCCGAATGGAC-3′ 5′-AGAGCCGAGAATAGGCACCC-3′ 121
CCNB1 5′-GAAGATCAACATGGCAGGCG-3′ 5′-GCATTTTGGCCTGCAGTTGT-3′ 131
BIRC3 5′-CCATTGACTTTTCTGTCGCCA-3′ 5′-GCATTATCCTTCGGTTCCCAA-3′ 125
GAPDH 5′-TGTTGCCATCAATGACCCCTT-3′ 5′-CTCCACGACGTACTCAGCG-3′ 201

Table 2

Clinicopathologic charateristics in HCC patients with or without liver-kidney yin deficiency syndrome(x±s)"

Group n Tumor size (cm) AFP level (μg/L) Age (years) γ-GT (U/L) ALT (U/L)
Liver-kidney yin
deficiency syndrome
3 3.77±0.14 165.67±95.99 57.33±1.20 165.67±95.99 47.33±14.77
Non-liver-kidney yin
deficiency syndrome
3 3.83±0.28 169.67±112.62 57.00±1.15 169.97±112.62 53.67±23.82

Figure 1

GO-MAP based on biological process for differentially expressed genes Red: significant GO category for down-regulated genes; Blue: significant GO category for up-regulated genes; Yellow: significant GO category for both up-regulated and down-regulated genes. GO: Gene Ontology."

Figure 2

KEGG pathway analysis for differentially expressed genes Blue: significant pathway for down-regulated genes; Red: significant pathway for up-regulated genes"

Figure 3

Interaction network analysis of differentially expressed genes Red: up-regulated genes; Blue: down-regulated genes"

Table 3

Genes with |Δ normalized degree|>0.3 in the interaction network analysis"

Gene Description ΔNormalized degree (D-C)
SEC62 SEC62 homolog
(S. cerevisiae)
–0.666 666 667
CCNB1 Cyclin B1 –0.372 549 020
BIRC3 Baculoviral IAP
repeat-containing 3
–0.341 176 471

Figure 4

The mRNA expressions of Sec62, Cyclin B1 and Birc3 in HCC patients with or without kidney-liver yin deficiency syndrome A: HCC patients without liver-kidney yin deficiency syndrome; B: HCC patients with liver-kidney yin deficiency syndrome. The Sec62/cyclin B1/Birc3 mRNA levels were expressed in comparsion with the expression level of GAPDH mRNA. Data were analyzed by Mann-Whitney U test, n=3; **P<0.01, vs group A. HCC: hepatocellular carcinoma; GAPDH: glyceraldehyde-3-phosphate dehydrogenase."

Figure 5

The protein expressions of Sec62, Cyclin B1 and Birc3 in HCC patients with or without kidney-liver yin deficiency syndrome A: HCC patients without liver-kidney yin deficiency syndrome; B: HCC patients with liver-kidney yin deficiency syndrome. The signal intensities were corrected with that of β-actin as an internal control. Data were analyzed by Mann-Whitney U test, n=3; **P<0.01, vs group A. HCC: hepatocellular carcinoma."

[1] Zhao XS, Luo R, Lei ZX, Xue YM Dong XL. Influence of genomics on modernization of traditional Chinese medicine syndrome. Yi Xue Yu Zhe Xue. 2003 ; 24(7): 57-58. Chinese.
赵晓山, 罗仁, 雷作熹, 薛耀明, 董晓雷 . 基因组学对中医证候现代化的影响.医学与哲学. 2003; 24(7):57-58.
[2] Hu B, An HM Shen KP. Liver-kidney yin deficiency is the basis of primary liver cancer. Zhongguo Zhong Yi Yao Xin Xi Za Zhi. 2009; 16(2) : 93. Chinese.
胡兵, 安红梅, 沈克平 . 肝肾阴虚是原发性肝癌的发病基础.中国中医药信息杂志. 2009; 16(2):93.
[3] Li HM, Li JP, Mao SS Liu L. Research on the distribution of syndrome of kidney-liver yin deficiency in liver diseases. Zhong Xi Yi Jie He Gan Bing Za Zhi. 2004; 14(5): 287-289. Chinese.
李瀚旻, 厉晶萍, 毛树松, 刘玲 . 肝病肝肾阴虚证型临床分布规律的研究.中西医结合肝病杂志. 2004; 14(5):287-289.
[4] Lin JH, Li HM, Li JP, Mao SS, Liu L . Observation and research on clinical features in patients with liver diseases of liver-kidney yin deficiency syndrome. Zhong Xi Yi Jie He Gan Bing Za Zhi. 2006; 16(4) : 239-241. Chinese.
林静华, 李瀚旻, 历晶萍, 毛树松, 刘玲 . 肝病患者肝肾阴虚证型临床相关特征的观察与研究.中西医结合肝病杂志. 2006; 16(4):239-241.
[5] Fang ZQ, Li YJ, Guan DY Chen DS. Suggestions about the standards of Chinese medical syndrome diferentiation for primary liver cancer. Shanghai Zhong Yi Yao Za Zhi. 2003; 37(5): 11-13. Chinese with abstract in English.
方肇勤, 李永健, 管冬元, 陈德溯 . 原发性肝癌中医辨证标准的建议.上海中医药杂志. 2003; 37(5):11-13.
[6] Zheng XY. Guidelines for clinical research on Chinese new herbal medicines. Beijing: China Press of Traditional Chinese Medicine. 2002: 4. Chinese.
郑筱萸 . 中药新药临床研究指导原则.北京: 中国中医药出版社. 2002: 4.
[7] Fang SQ, Lin BH, Ye Y, Wu SX, Chen K, Lin QC . Study on the relationship of kidney deficiency syndrome, hormone and immunity in old people. Fujian Zhong Yi Yao. 2002; 33(2): 1-3. Chinese.
方素钦, 林炳辉, 叶盈, 吴圣贤, 陈可, 林求诚 . 中老年人肾虚证与性激素及免疫功能的研究.福建中医药. 2002; 33(2) : 1-3.
[8] Quan JF, Wu XK, Sun XH . Association of serum immunoglobulin G, A and M and complement C3 and C4 in patients with kidney deficiency syndrome. Xian Dai Zhong Yi Yao. 2004; 5(5): 53-54. Chinese.
全建峰, 吴晓康, 孙晓红 . 肾阴虚证患者的血清免疫球蛋白G、A、M及补体C3、C4相关性研究.现代中医药. 2004; 5(5) : 53-54.
[9] Guo JR, Yan HF . Correlation between deformability of red blood cell in saliva and kidney yin deficiency syndrome in chronic nephritis. Zhong Yi Yao Xue Kan. 2003; 21(11): 1900-1901. Chinese.
郭金瑞, 严惠芳 . 慢性肾炎唾液Na+、K+红细胞变形能力改变与肾阴虚证相关性研究.中医药学刊. 2003; 21(11):1900-1901.
[10] Gao Y, Gao H, Wang MQ . Analysis of differentially expressed genes in a diabetes family of dual deficiency of kidney yin and kidney yang with blood stasis. Xian Dai Zhong Xi Yi Jie He Za Zhi. 2005; 14(1): 1-5. Chinese.
高黝, 高泓, 王米渠 . 1个糖尿病家系肾阴阳两虚血瘀证糖尿病的差异表达基因分析.现代中西医结合杂志. 2005; 14(1) : 1-5.
[11] Zhong YF, Chen YP, Deng YY, Chen N, Wang ZH . Correlation between traditional Chinese medicine syndromes in primary immunoglobulin A nephropathy and A267G in 5’-untranslated region within exonal of megsin gene. J Chin Integr Med. 2008; 6(5): 463-467. Chinese with abstract in English.
钟逸斐, 陈以平, 邓跃毅, 陈楠, 王朝晖 . Megsin基因El-5'UTR区A267G与免疫球蛋白A型肾病阴虚证的相关性.中西医结合学报. 2008; 6(5):463-467.
[12] Su W, Xu ZY . Gene polymorphism and physical constitution theory : starting point of exploring syndromes of lung cancer. J Chin Integr Med. 2010; 8(1): 15-19. Chinese with abstract in English.
苏婉, 徐振晔 . 从基因多态性及体质学说探讨肺癌的中医证候.中西医结合学报. 2010; 8(1):15-19.
[13] Schlitt T, Palin K, Rung J, Dietmann S, Lappe M, Ukkonen E, Brazma A . From gene networks to gene function. Genome Res. 2003 ; 13(12) : 2568-2576.
doi: 10.1101/gr.1111403
[14] Yi M, Horton JD, Cohen JC, Hobbs HH, Stephens RM . WholePathwayScope : a comprehensive pathway- based analysis tool for high-throughput data. BMC Bioinformatics. 2006; 7:30.
doi: 10.1186/1471-2105-7-30
[15] Pujana MA, Han JD, Starita LM, Stevens KN, Tewari M, Ahn JS, Rennert G, Moreno V, Kirchhoff T, Gold B, Assmann V, Elshamy WM, Rual JF, Levine D, Rozek LS, Gelman RS, Gunsalus KC, Greenberg RA, Sobhian B, Bertin N, Venkatesan K, Ayivi-Guedehoussou N, Sole X, Hernandez P, Lazaro C, Nathanson KL, Weber BL, Cusick ME, Hill DE, Offit K, Livingston DM, Gruber SB, Parvin JD, Vidal M . Network modeling links breast cancer susceptibility and centrosome dysfunction. Nat Genet. 2007; 39(11):1338-1349.
doi: 10.1038/ng.2007.2
[16] Ai H, Niu JZ, Xue XO, Li J . Effects of genistein on ovary gene expression pattern of CDDP induced premature ovarian failure mice with syndrome of kidney and liver yin asthenia. Beijing Zhong Yi Yao Da Xue Xue Bao. 2008; 31(7): 452-455. Chinese with abstract in English.
艾浩, 牛建昭, 薛晓鸥, 李健 . 金雀异黄素对肝肾阴虚证卵巢功能早衰小鼠基因差异表达作用研究.北京中医药大学学报. 2008; 31(7):452-455.
[17] Wei M, Zhao XS, Sun XM Luo R. Differential gene expression in kidney-yin deficiency. Guangdong Yi Xue. 2010; 31(23) : 3017-3020. Chinese with abstract in English.
魏敏, 赵晓山, 孙晓敏, 罗仁 . 肾阴虚证患者的基因差异表达.广东医学. 2010; 31(23):3017-3020.
[18] Panzner S, Dreier L, Hartmann E, Kostka S, Rapoport TA . Posttranslational protein transport in yeast reconstituted with a purified complex of Sec proteins and Kar2p. Cell. 1995; 81(4) : 561-570.
doi: 10.1016/0092-8674(95)90077-2
[19] Tyedmers J, Lerner M, Bies C, Dudek J, Skowronek MH, Haas IG, Heim N, Nastainczyk W, Volkmer J, Zimmermann R . Homologs of the yeast Sec complex subunits Sec62p and Sec63p are abundant proteins in dog pancreas microsomes. Proc Natl Acad Sci USA. 2000; 97(13) : 7214-7219.
doi: 10.1073/pnas.97.13.7214
[20] Dudek J, Greiner M, Muller A, Hendershot LM, Kopsch K, Nastainczyk W, Zimmermann R . ERjlp has a basic role in protein biogenesis at the endoplasmic reticulum. Nat Struct Mol Biol. 2005; 12(11):1008-1014.
doi: 10.1038/nsmb1007
[21] Muller L, de Escauriaza MD, Lajoie P, Theis M, Jung M, Muller A, Burgard C, Greiner M, Snapp EL, Dudek J, Zimmermann R . Evolutionary gain of function for the ER membrane protein Sec62 from yeast to humans. Mol Biol Cell. 2010; 21(5):691-703.
doi: 10.1091/mbc.e09-08-0730
[22] Greiner M, Kreutzer B, Jung V, Grobholz R, Hasenfus A, Stohr RF, Tornillo L, Dudek J, Stockle M, Unteregger G, Kamradt J, Wullich B, Zimmermann R . Silencing of the SEC62 gene inhibits migratory and invasive potential of various tumor cells. Int J Cancer. 2011 ; 128(10):2284-2295
doi: 10.1002/ijc.25580
[23] Nimeus-Malmstrom E, Koliadi A, Ahlin C, Holmqvist M, Holmberg L, Amini RM, Jirstrom K, Warnberg F, Blomqvist C, Ferno M, Fjallskog ML . Cyclin B1 is a prognostic proliferation marker with a high reproducibility in a population-based lymph node negative breast cancer cohort. Int J Cancer. 2010; 127(4):961-967.
[24] Murakami H, Furihata M, Ohtsuki Y, Ogoshi S . Determination of the prognostic significance of cyclin B1 overexpression in patients with esophageal squamous cell carcinoma. Virchows Arch. 1999 ; 434(2) : 153-158.
doi: 10.1007/s004280050319
[25] Soria JC, Jang SJ, Khuri FR, Hassan K, Liu D, Hong WK, Mao L . Overexpression of cyclin B1 in early-stage non-small cell lung cancer and its clinical implication. Cancer Res. 2000; 60(15):4000-4004.
[26] Begnami MD, Fregnani JH, Nonogaki S, Soares FA . Evaluation of cell cycle protein expression in gastric cancer : cyclin B1 expression and its prognostic implication. Hum Pathol. 2010; 41(8):1120-1127.
doi: 10.1016/j.humpath.2010.01.007
[27] Calvisi DF, Simile MM, Ladu S, Frau M, Evert M, Tomasi ML, Demartis MI, Daino L, Seddaiu MA, Brozzetti S, Feo F, Pascale RM . Activation of v-Myb avian myeloblastosis viral oncogene homolog-like2 (MYBL2 ) -LIN9 complex contributes to human hepato- carcinogenesis and identifies a subset of hepatocellular carcinoma with mutant p53. Hepatology. 2011; 53(4):1226-1236.
doi: 10.1002/hep.24174
[28] Nachmias B, Ashhab Y, Ben-Yehuda D . The inhibitor of apoptosis protein family (IAPs) : an emerging therapeutic target in cancer. Semin Cancer Biol. 2004; 14(4) : 231-243.
doi: 10.1016/j.semcancer.2004.04.002
[29] Yang D, Yaguchi T, Nakano T, Nishizaki T . Adenosine- induced caspase-3 activation by tuning Bel-XL/DIABLO/ IAP expression in HuH-7 human hepatoma cells. Cell Biol Toxicol. 2010; 26(4):319-330.
doi: 10.1007/s10565-009-9145-7
[30] Du WX, Zhu MD, Jiang M, Ma P Zhou Y. Research on the essence of traditional Chinese medicine syndrome under the guidance of biological systematology. Zhongguo Zhong Xi Yi Jie He Za Zhi. 2011; 31(3): 419-423. Chinese.
杜武勋, 朱明丹, 姜民, 马朋, 周莹 . 生物系统论指导下的中医证候实质研究及其问题.中国中西医结合杂志. 2011; 31(3):419-423.
[31] Zhang DH, Ling CQ, Liu Y, Ye ZB, Bao YX, Kong XT . Modern cytobiology and traditional Chinese medicine theory : the relationship between Bcl-2 family and the essence of kidney in traditional Chinese medicine. Xi Bao Sheng Wu Xue Za Zhi. 1999; 21(3): 125-128. Chinese.
张登海, 凌昌全, 刘颖, 叶志斌, 鲍一笑, 孔宪涛 . 现代细胞生物学与中医学说——Bcl-2家族与中医肾本质关系探讨.细胞生物学杂志. 1999; 21(3):125-128.
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