The Study On ALDH1 As A Marker Of Cervical Cancer Stem Cells

[Objective]To investigate and verify the possibility of ALDH1 being the markers of cervical cancer, and the possibility of jointing screening CCSCs by SP and ALDH1.[Methods]1. Obtain ALDH1 high and ALDH1 low cells respectively, by Aldefluor fluorescent staining of cervical cancer cell lines Si Ha and cervical cancer primary cells.2. Observing the self-renewal ability of ALDH1 high and ALDH1 low cells by serum-free suspension culture experiment.3. Observing the proliferation ability of ALDH1 high and ALDH1 low cells by soft AGAR cloning experiment.4. Using reverse transcription- polymerase chain reaction(Real- Time PCR) to detect the stem cell gene expression of ALDH1 high and ALDH1 low cells.5. Determining the relationship between the two cell subsets by jointing screening CCSC by SP and ALDH1.[Results]1. The ALDH1 high cells were Detected by flow cytometry and ALDH1 high cells were(0.77±0.25)% in cervical cancer cell lines Si Ha(n=6),(3.13±0.85)% in cervical cancer primary cells(n=6).2. The results of the serum-free suspension culture experiment: the sphere formation rate of ALDH1 high cells were(12.91±3.94)% which is higher than ALDH1 low cells(5.83±0.96)% in cervical cancer cell lines Si Ha,(n=4, P <0.05); the sphere formation rate of ALDH1 high cells were(22.92±2.84)% which is higher than ALDH1 low cells(8.33±4.90)% in cervical cancer primary cells,(n=4, P <0.05).3. The number of cell clone formation in soft AGAR: The number of cell clone formation of ALDH1 high cells were(26.17±7.63)/200 cells which is higher than ALDH1 low cells(12.83±2.56)/200 cells in cervical cancer cell lines Si Ha,(n=6, P<0.05); The number of cell clone formation of ALDH1 high cells were(55.00±8.67)/200 cells which is higher than ALDH1 low cells(23.67±3.93)/200 cells in cervical cancer primary cells,(n=6, P <0.05).4. The gene expression of ALDH1 high and ALDH1 low cells in cervical cancer primary cells: the expression of CD24 and CD44 in ALDH1 high cells is significantly higher than in ALDH1 low cells, is(2.23±0.33) times and(1.87±0.12) times respectively(n=3, P <0.05); the expression of Nanog and Oct4 in ALDH1 high cells is slightly higher than in ALDH1 low cells, is(1.41±0.27) times and(1.49±0.20) times respectively(n=3, P >0.05).5. The proportion of ALDH1 high cells in SP cells were 0.75%, and the proportion of ALDH1 low cells in SP cells were 5.87%. SP and ALDH1 high cell subsets do not overlap.[Conclusions]1. The self-renewal and proliferation abilities of ALDH1 high cells were higher than ALDH1 low cells cervical cancer cell lines Si Ha and cervical cancer primary cells.2. The the stem cell gene expression of ALDH1 high cells were higher than ALDH1 low cells cervical cancer cell lines Si Ha and cervical cancer primary cells.3. CCSCs can be enriched by ALDH1, but the purity is not enough, it can’t be a single marker of CCSCs purification.4. There is little possibility of jointing screening CCSCs by SP and ALDH1.