A pilot study of differential genes in nonsyndromic cleft lip with or without cleft palate（NSCL/P） by expression microarray

Abstract:

Objective：To analyze the deferentially expressed genes between nonsyndromic cleft lip with or without cleft palate（NSCL/P） patients and healthy control using gene expression microarray technology and to screen NSCL/P related genes. Methods：Total RNA was extracted from 3 NSCL/P patients and 3 controls by centrifugal column method. The RNA was purified and qualitatively qual－ified，then reverse transcribed into cDNA. The cDNA was transcribed into cRNA；fluorescence dye Cy3 was used to label the cRNA. The labeled cRNA was used to hybrid with the Agilent 4 × 44 k human genome-wide expression of the gene chip；the fluorescence signal was scanned by the computer and the original data were normalized. The differential expression genes was screened by t-test and fold difference.The gene functional annotation and correlation analysis were carried out by the online analysis system of Shanghai Bohao biology to clarify the biological functions of the different genes. The specific differential expression genes were selected and the reliability of the chip results was verified by real-time PCR. Results：Totally 254 differentially expressed genes were screened out，among which 151 had up-regulated expression and 103 had down-regulated expression. Five differential expression genes were selected for real-time PCR. ADH1C（t=5.132，P=0.000），RDH10（t=2.960，P=0.039），HIST1H2BD（t=4.446，P=0.001） had down-regu－lated expression，KAT2B（t=-4.945，P=0.000），FOSB（t=-3.666，P=0.005） had up-regulated expression. The results showed that three genes had down-regulated expression and two genes had up-regulated expression，which was consistent with the result of chip ex－pression. Conclusion：There are significant differences in gene expression between normal children and children with NSCL/P. Analyzing these diferentially expressed genes may help clarify the pathogenesis of NSCL/P and provide a theoretical basis for disease prevention.