[Genetic instability detected by flow cytometry: DNA aneuploid and P16 expression in biopsy specimens from lung cancer].

OBJECTIVE: To investigate DNA aneuploid and P16 expression in biopsy specimens from lung cancer, and to study genetic instability and the application of flow cytometry in lung cancer pernicious degree diagnosis. METHODS: Blood cells and cancer cells in biopsy specimens were marked simultaneously with anti-CD45 and anti-P16 fluorescent antibody, and the ratio of CD45+ P16+ cells and CD4- P16+ cells was compared. DNA content in biopsy specimens from lung cancer was detected by flow cytometry. RESULTS: Among the 74 cases of lung cancer, there are 46 cases of DNA aneuploid (62.2%). Thirty-seven cases of lung cancer expressed P16 lowly (50%). Twelve cases of lung cancer only expressed P16 lowly (16.22%), 21 cases of lung cancer only expressed DNA aneuploid (28.38%), and 25 cases not only expressed P16 lowly but also expressed DNA aneuploid (33.78%). Indexes of malign degree, such as P16 low expression or DNA aneuploid could be detected in 58 cases among the 74 cases (78.38%) by flow cytometry. CONCLUSION: P16 low expression and DNA aneuploid are the indexes of lung cancer malign degree, and flow cytometry can be used to study genetic instability and evaluate biopsy specimens from lung cancer.

[The expression of mutation p53 gene from circulating cancer cells of peripheral blood and the clinical significance in detecting the patients with colorectal cancer].

OBJECTIVE: To study the clinical significance of detecting p53 gene mutation expression in colorectal cancer cells of peripheral blood. METHODS: Flow cytometry (FCM) was used to detect p53 gene mutation expression in peripheral blood cancer cells of 128 patients with colorectal cancer. Experimental data were analyzed by SPSS (v.11.0) software. RESULTS: The lymph node metastasis showed the significant difference statistically (P<0.01) between p53 positive and negative expression in the colorectal cancer patients. The mutation p53 expression associated with existing histological differentiation (r=0.8476, P<0.05). A lymph node metastasis difference was observed between left and right colorectal cancers of mutation p53 positive expression. CONCLUSION: Detecting the mutation p53 expression in cancer cells of peripheral blood might be helpful to the early diagnosis of colorectal cancer.

[Effect of IL-6/sIL-6R on ex vivo expansion of human cord blood derived CD34+ cells].

BACKGROUND & OBJECTIVE: Hematopoietic stem cells (HSC) have the ability of regeneration, differentiation, and reconstructing hematopoietic function, and it is widely used in many fields such as hematopoietic stem cell transplantation, immune therapy, gene therapy and so on. Human cord blood (CB) is abundant of HSC. But a single collection of CB has only a limited amount of HCS and cannot fit the clinical and research use. Thus ex vivo expansion of human CB derived HSC is important. We know that there are some cytokines, which can synergize for enhancing the expansion of CB derived CD34(+) cells in vitro. Currently, some experiments have discovered that IL-6/sIL-6R or its chimera can enhance the ex vivo expansion of CD34(+)gp130+IL-6R- subpopulation. This study was designed to observe the effect of IL-6/sIL-6R on the ex vivo expansion of human CB derived CD34(+) cells, and explore the optimal cytokine combinations. METHODS: Human CB derived CD34(+) cells were isolated by Mini MACS and cultured in ex vivo liquid media in the presence of different cytokine cocktails for 7 or 14 days. After cultured on the seventh or the fourteenth day, the total number of the cultured cells were counted, the ratio of the CD34(+) cell were assayed by flow cytometry (FCM) and the number of it were calculated, and CFU-GM were cultured, then the effects of different cytokine combinations on the ex vivo expansion of CD34(+) cells were compared. In line with the different cytokine cocktails, our experiment divided into five groups: (A) control,(B) SCF,(C) IL-6/sIL-6R+SCF,(D) IL-6/sIL-6R+SCF+FL,and (E) SCF+FL. RESULTS: After cultured in vitro for 7 or 14 days, (1) the number of CD34(+) cells descended apparently in groups A and B; (2) the number of nucleated cells and CD34(+) cells after cultural on the seventh or the fourteenth day increased 7.1+/-2.4 folds, 39.0+/-14.0 folds; 1.8+/-0.7 folds, 4.8+/-2.4 folds, respectively in group C; 16.5+/-5.7 folds, 110.0+/-28.0 folds; 3.5+/-1.5 folds, 10.2+/-4.2 folds in Group D; 17.3+/-3.8 folds, 104.0+/-21.0 folds; 3.6+/-2.1folds, 8.4+/-3.5 folds in Group E. The expansion effects of group C, D, and E were all superior to the group A or B (P< 0.01). The expansion effects of group D and E were superior to group C (P< 0.01). But there was no difference between group D and E (P >0.05); (3) Adding the concentration of sIL-6R to 400 ng/ml, the number of nucleated and CD34(+) cells increased 24.0+/-4.8 folds and 5.6+/-1.2 folds in group D after cultured for seven days superior to group E (P< 0.05). CONCLUSION: IL-6/sIL-6R, SCF, FL can synergize for enhancing the ex vivo expansion of human CB derived CD34(+) cells. But this synergetic effect depends on the concentration of sIL-6R.