Supplementary data
Figure legends
Supplementary Figure 1 Laser capture microdissection of tumor sample from patient 397. Laser capture microdissection was performed to obtain tumor cells from H&E-stained lung tumor section of patient 397. The histology of the H&E-stained section from paraffin embedded tumor sample was shown in A (left panel, 100X; right panel, 400X). The change of a section before and after microdissection is shown in B and C.
Supplementary Figure 2 Sequencing results for LKB1 mutations from lung
adenocarcinoma and normal lung samples. Genomic DNA from lung
adenocarcinoma and corresponding distant normal lung samples were sequenced for
LKB1 mutations. The five patients listed here have heterozygous mutations in their
lung tumor samples, which are absent in the paired normal lung samples.
Supplementary Tables
Supplementary Table 1 Primers for PCR and DNA sequencing to analyze LKB1 mutations in Chinese lung adenocarcinomas
PCR primers Sequencing primers Exon 1F 5’ ggactgacgtgtagaacaatcg3’
Exon 1R 5’ ctggccttgcctgagtgaaagt3’
Exon 1R 5’ ctggccttgcctgagtgaaagt3’
Exon 2F 5’ gaggtacgccacttccacag3’
Exon 2R 5’ cttcaaggagacgggaagag3’
Exon 2F2 5’ agggatccaggccatcat3’
Exon 3F 5’ aagagtcagccctgtcct3’
Exon 3R 5’ atcaggacaagcagtgtgg3’
Exon 3F2 5’ ggcaaggtgggtgcagag3’
Exon 4-5F 5’ cctggacttctgtgacttcc3’
Exon 4-5R 5’ gtgaaccaccatctgccgtat3’
Exon 4-5F2 5’ agctgggcctgtggtgtttg3’
Exon 6F 5’ cactagtggaaggtggtgaaga3’
Exon 6R 5’ tctccactcagtcctctcaatg3’
Exon 6R2 5’ acccccaaccctacatttct3’
Exon 7F 5’ ttaggagcgtccaggtatca3’
Exon 7R 5’ taacaggacactgcccagaga3’
Exon 7R2 5’ ctcaaccagctgcccacat3’
Exon 8F 5’ ctgagcttctgtggtcacag3’
Exon 8R 5’ acgtgggattggccaccaga3’
Exon 8F2 5’ tcctgagtgtgtggcaggta3’
Exon 9F 5’ tgaggttcctgcagtcagtac3’
Exon 9R 5’ tgacggtcaccatgactgacta3’
Exon 9F2 5’ gtaagtgcgtccccgtggtg3’
Supplementary Table 2 PCR and sequencing primers for EGFR and KRAS mutation detection using cDNA and/or genomic DNA
Primers for cDNA PCR and sequencing
Gene PCR primers Sequencing primers
EGFR
F1 5’ caacaccctggtctggaagt 3’
R1 5’ aggcgttctcctttctccag 3’
F2 5’ ggctgctgcaggagagggag 3’
R2 5’ aggtcatcaactcccaaacg3’
KRAS
F1 5’ggagagaggcctgctgaaa 3’
R1 5’tcctgagcctgttttgtgtct 3’
R2 5’ctgttctagaaggcaaatcaca 3’
Primers for genomic DNA PCR and sequencing
Gene PCR primers Sequencing primers
EGFR
Exon 18F 5’agcatggtgagggctgaggtgac3’
Exon 18R 5’atatacagcttgcaaggactctgg3’
Exon 18R
5’atatacagcttgcaaggactctgg3’
Exon 19F 5’agatcactgggcagcatgtg3’
Exon 19R 5’atgagcagggtctagagcaga3’
Exon 19R 5’atgagcagggtctagagcaga3’
Exon 20F 5’ gatcgcattcatgcgtcttcacc3’
Exon 20R 5’ ttgctatcccaggagcgcagacc3’
Exon 20F 5’ gatcgcattcatgcgtcttcacc3’
Exon 21F 5’ tcagagcctggcatgaacatgac3’
Exon 21R 5’ ggtccctggtgtcaggaaaatg3’
Exon 21F 5’ tcagagcctggcatgaacatgac3’
KRAS
Exon 2F 5’ taagcgtcgatggaggagttt3’
Exon 2R 5’ ccctgacatactcccaaggaa3’
Exon 2R 5’ccctgacatactcccaaggaa3’
Supplementary Table 3 Tumor cell content statistics of 86 Chinese lung adenocarcinomas
Patient ID
Pathological analyses of small tumor chunks
# 1 # 2 # 3 # 4
TC (%)
FB (%)
IC (%)
TC (%)
FB (%)
IC (%)
TC (%)
FB (%)
IC (%)
TC (%)
FB (%)
IC (%) 97 70 20 10 70 20 10 70 30 0 50 50 0 105 60 40 0 70 30 0 50 50 0 70 10 20 106 70 10 20 80 10 10 70 20 10 60 30 10 116 70 20 10 70 20 10 70 20 10 50 40 10 123 70 20 10 40 40 20 70 10 20 40 30 30 128 70 30 0 70 20 10 70 20 10 70 15 15 129 70 15 15 30 60 10 20 40 40 70 20 10 136 20 60 20 80 15 5 70 20 10 80 15 5 154 70 30 0 60 40 0 30 65 5 70 15 15 163 70 25 5 40 40 20 30 60 10 70 20 10 168 80 15 5 30 60 10 50 45 5 70 25 5 175 70 20 10 50 40 10 70 20 10 60 20 20 178 70 20 10 50 40 10 70 25 5 70 25 5 180 70 10 20 70 15 15 70 10 20 40 50 10 181 70 20 10 70 15 15 60 10 30 50 30 20 182 70 25 5 70 15 15 70 10 20 70 15 15 187 50 30 20 50 25 25 70 10 20 70 10 20 194 70 25 5 40 50 10 70 20 10 60 30 10 197 70 15 15 70 20 10 70 20 10 70 15 15 198 70 25 5 80 15 5 60 35 5 60 35 5 202 80 15 5 80 20 0 80 20 0 80 15 5 203 70 20 10 60 10 30 70 20 10 70 25 5 212 80 20 0 70 30 0 70 20 10 80 20 0 218 80 15 5 80 10 10 80 15 5 80 15 5 224 70 10 20 70 0 30 70 0 30 80 0 20 230 80 15 5 70 20 10 70 10 20 70 20 10 231 80 0 20 70 10 20 70 10 20 40 50 10 240 70 10 20 50 30 20 20 50 30 70 25 5 243 60 35 5 70 25 5 70 20 10 70 10 20 245 70 10 20 70 15 15 70 20 10 60 30 10 249 80 10 10 70 10 20 70 25 5 70 25 5 251 70 30 0 70 30 0 70 30 0 60 35 5 259 10 30 60 70 20 10 30 50 20 70 20 10 262 70 25 5 70 30 0 80 10 10 80 20 0 264 60 30 10 20 70 10 80 5 15 70 10 20
267 80 20 0 70 25 5 70 15 15 60 35 5 269 70 20 10 70 15 15 70 10 20 70 15 15 272 70 25 5 70 15 15 50 40 10 70 20 10 275 70 20 10 70 25 5 70 25 5 70 25 5 277 60 35 5 70 30 0 70 25 5 50 40 10 278 70 25 5 60 35 5 60 35 5 70 25 5 280 80 15 5 70 5 25 80 5 15 80 5 15 291 80 15 5 70 25 5 70 25 5 70 25 5 300 70 20 10 80 15 5 70 25 5 70 20 10 305 50 40 10 50 10 40 70 25 5 70 25 5 309 50 40 10 70 5 25 80 10 10 60 30 10 316 80 15 5 70 25 5 60 30 10 70 25 5 318 60 30 10 70 20 10 70 20 10 50 40 10 324 60 30 10 70 10 20 70 20 10 10 40 50 325 70 20 10 70 20 10 70 20 10 60 10 30 327 60 30 10 70 20 10 70 20 10 80 10 10 329 40 50 10 70 20 10 70 20 10 50 40 10 336 70 25 5 70 25 5 70 25 5 80 20 0 338 70 0 30 60 10 30 70 30 0 30 50 20 339 80 15 5 80 15 5 70 30 0 50 40 10 345 80 0 20 80 10 10 80 10 10 80 10 10 346 70 20 10 70 20 10 70 25 5
347 80 20 0 60 30 10 80 15 5 70 20 10 350 70 10 20 70 10 20 60 10 30 70 5 25 355 80 10 10 70 20 10 60 30 10 50 40 10 363 30 60 10 70 25 5 70 25 5 70 20 10 368 70 25 5 80 15 5 70 15 15 60 25 15 370 50 40 10 70 25 5 70 20 10 30 50 20 373 40 30 30 70 5 25 70 5 25 50 5 45 374 30 60 10 70 20 10 70 20 10
380 50 10 40 70 10 20 70 10 20 60 20 20 382 70 0 30 70 5 25 20 20 60
385 70 25 5 80 10 10 80 10 10 80 5 15 392 70 10 20 50 40 10 70 20 10 50 40 10 393 80 15 5 70 20 10 70 25 5 70 25 5 397 80 15 5 70 20 10 80 10 10 70 10 20 399 70 20 10 70 20 10 70 15 15 70 20 10 411 70 25 5 70 20 10 60 30 10 70 20 10 412 80 15 5 80 20 0 80 10 10 70 20 10 421 70 10 20 70 5 25 70 20 10
423 50 10 40 70 20 10 70 15 15 80 5 15 435 50 10 40 70 10 20 70 15 15 60 10 30
448 80 10 10 80 10 10 70 20 10 80 5 15 456 70 20 10 70 20 10 70 20 10 80 20 0 481 80 0 20 70 0 30 70 0 30 60 10 30 485 10 30 60 50 15 35 70 5 25 70 10 20 491 70 25 5 70 25 5 50 40 10 60 35 5 511 70 25 5 80 20 0 70 25 5 40 10 50 512 70 15 15 70 20 10 70 20 10 40 50 10 513 5 85 10 80 10 10 80 15 5 30 60 10 TC: tumor cell; FB: fibroblasts; IC: inflammatory cells.
