Table 3. Cyclin D2 and PTCH1 expression and Promoter hypermethylation analysis in astrocytic cell lines

Astrocytic Cell lines / Cyclin D2 expression / Cyclin D2
Expression after 5’aza+TSA / Cyclin D2 promoter hypermethylation analysis by MCA- MSP / Cyclin D2 promoter hypermethylation
Analysis by MCA-Meth / Cyclin D2 Hypermethylation by both techniques / No Hypermethylation in Cyclin D2 promoter / PTCH1 expression in glioma cell lines / PTCH1 promoter hypermethylation by MCA-MSP
U87MG / + / + / Ö / Ö / Ö / +/-
A172 / - / + / Ö / -
LN405 / + / + / Ö / +
SW1783 / - / + / Ö / -
T98G / - / ++ / Ö / Ö / Ö / ++
SW1088 / + / + / Ö / ++
CCF-STTG-1 / - / + / Ö / ++ / Ö
GOS-3 / - / ++ / Ö / Ö / Ö / ++

-: no expression; +: expression; ++: high expression; Ö: yes

Table 3. Promoter methylation and expression correlation among eight astrocytic cell lines of Cyclin D2 and PTCH1 gene. Table shows that 63% (5/8) of cell lines have hypermethylation in the Cyclin D2 promoter region and absence of Cyclin D2 transcript expression. In PTCH1 gene 13% (1/8) of the cell line shows promoter hypermethylation but there is not any correlation with hypermethylation in the promoter region of PTCH1.

Table 4. Expression and promoter hypermethylation analysis of Cyclin D2 and PTCH1 gene in astrocytic tumor samples

Astrocytic tumor samples Grade
and Sample ID / Expression of Cyclin D2 in Samples / Methylation by MSP in Cyclin D2 promoter / Methylation by Melting curve in Cyclin D2 promoter / Methylation by both techniques in Cyclin D2 promoter / No hypermethylation in Cyclin D2 promoter / Expression of PTCH1 transcript in tumor
samples / Methylation of PTCH1 promoter by MCA-Meth
GBM(3) / - / Ö / Ö / Ö / - / Ö
AIII(4) / ++ / Ö / +
GBM(5) / - / Ö / +/-
GBM(6) / - / Ö / +/-
AIII(7) / + / Ö / +/-
GBM(8) / - / Ö / ++
GBM(9) / + / Ö / -
AIII(10) / - / Ö / +/-
GBM(11) / ++ / Ö / +/-
GBM(12) / + / Ö / +
GBM(13) / - / Ö / Ö / Ö / + / Ö
AIII(14) / ++ / Ö / -
GBM(15) / - / Ö / +/-
AIII(16) / ++ / Ö / +/-
GBM(17) / - / Ö / ++
GBM(18) / - / Ö / Ö / Ö / +
AIII(19) / + / Ö / +
AII(20) / + / Ö / +
AIII(21) / - / Ö / - / Ö
GBM(22) / - / Ö / -
GBM(23) / - / Ö / +
GBM(24) / - / Ö / Ö / Ö / +
AI(25) / ND / Ö / ND
GBM(26) / ND / Ö / ND
GBM(27) / ND / Ö / ND
GBM(28) / ND / Ö / ND
GBM(29) / ND / Ö / ND
GBM(30) / + / Ö / +
GBM(31) / ND / Ö / ND
GBM(32) / ND / Ö / ND
AI(34) / ND / Ö / ND
AIII(35) / ND / Ö / ND
GBM(36) / ND / Ö / ND
GBM(41) / ND / Ö / ND
GBM(43) / + / Ö / -
GBM(44) / ND / Ö / ND
GBM(45) / ND / Ö / ND
GBM(46) / ND / Ö / ND
GBM(47) / ND / Ö / ND
GBM(48) / ND / Ö / ND
GBM(49) / ND / Ö / ND
AII(50) / ++ / Ö / -
GBM(51) / - / Ö / -
GBM(52) / ++ / Ö / +

-:no expression; +/-: low expression; +: expression; ++: high expression; Ö:yes; nd: not determined

Table 4. Promoter hypermethylation and transcript expression correlation of Cyclin D2 and PTCH1 gene among primary astrocytic tumor samples. Table.2 shows 93% (13/14) of samples do not express Cyclin D2 transcript and have hypermethylation in the Cyclin D2 promoter region. Only 67% (2/3) of the samples show correlation with absence of PTCH1 transcript expression and promoter hypermethylation in PTCH1 gene.