Table 1.
Expression of Viral Genes and Cellular Targets was Analyzed in 6 Cell Lines, 3 Normal Cervix and 10 Clinical Samples in Relation to HPV DNA Status
| HPV16-Samples | E2-E4 | E2N | E6-E7 | Ratio E2N/E6-E7 | Ratio E2E4/E6-E7 | CyclinB1 | E2 IHC | P16INK4 | HPV DNA |
|---|---|---|---|---|---|---|---|---|---|
| Caski | 11000 | 2500 | 8500 | 0.3 | 1.3 | 4600 | - | ND | Int |
| Caski+E2 | 340 | 200 | 300 | 0.6 | 1.1 | 390 | - | ND | Int |
| SiHa | 4 | 360 | 1930 | 0.18 | - | 3900 | - | ND | Int |
| IC3 | 5 | 1960 | 4900 | 0.4 | - | 2300 | - | ND | Int |
| C33 | 4 | 5 | 2 | - | - | 3300 | - | ND | No |
| N-TERT | 5 | 7 | 4 | - | - | 2700 | - | ND | No |
| 2Ncervix | 5 | 9 | 2 | - | - | 130 | - | - | No |
| 4Ncervix | 5 | 10 | 6 | - | - | 110 | - | - | No |
| 8Ncervix | 4 | 4 | 3 | - | - | 60 | - | - | No |
| 2CIN2 | 15800 | 1520 | 570 | 2.6 | 27 | 290 | +++ | ++ | Rep ++ |
| 3CIN2 | 21500 | 4000 | 3100 | 1.3 | 7 | 1060 | +++ | ++ | Rep ++ |
| 4CIN2 | 33000 | 3900 | 2300 | 1.7 | 14 | 870 | +++ | + | Rep +++ |
| 2CIN3 | 3600 | 520 | 1130 | 0.5 | 3 | 460 | - | +++ | Rep+/- Int? |
| 12CIN3 | 136 | 29 | 70 | 0.4 | 2 | 200 | +/- | + | Rep+/- Int? |
| 18CIN3 | 81 | 38 | 38 | 1 | 2 | 190 | - | +++ | Rep+/- Int? |
| 2SCC | 3090 | 860 | 1680 | 0.5 | 1.8 | 600 | + | ++ | Int + rep? |
| 6SCC | 73600 | 37000 | 31500 | 1.17 | 2.3 | 3200 | - | +++ | Int |
| 7SCC | 11600 | 1500 | 5330 | 0.28 | 2.2 | 2000 | +/- | ++ | Int+ rep? |
| 8SCC | 15200 | 2200 | 7800 | 0.28 | 1.9 | 1600 | - | +++ | Int |
Transcription of the viral and cellular genes was measured by digital quantification using the NanoString technology. For each probe, the table shows the actual number of RNA molecules to which the probe has been bound, after standardization with 3 housekeeping genes: RPL27, RPS13 and ACTB. Coefficients of variation are typically < 7% for values above 10. Protein expression of E2 and p16INK4 was determined by immunohistochemistry (IHC) and HPV DNA status was assessed by in situ hybridization (ISH). RNA extraction, ISH and IHC were performed on serial sections of the same samples. Staining was summarized as +++ when at least half, ++ one third and + less than 1/3 of the cells of the samples were stained. For ISH, the 2 types of signals are indicated as replication (Rep) for uniform staining of the nuclei or integrated (Int) for homogenous punctate staining as shown in Fig. (3B). ND: Not Determined.