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Abstract
Purpose
This study investigated the inhibitory effects of Paclitaxel by altering tubulin assembly and cisplatin exposure by binding DNA of the lens epithelial cells (LECs) during epithelial cell cultures in the capsular bag model.
Methods
In the capsular bag model, the LECs were cultured with exposure to Paclitaxel (1, 10, 100 nM) and Cisplatin (1, 10, 100 µM) for 3 min. The effect of Paclitaxel and Cisplatin was analyzed by observing the cell number of fibroblasts per field, Western blots for type IV collagen, TUNEL assay and the Proliferating Cell Nuclear Antigen (PCNA) and Bromodeoxyuridine (BrdU) incorporated proliferating cells.
Results
An increase in concentration of Paclitaxel and Cisplatin resulted in a decrease in the number of fibroblasts and spindle-shaped cells. The number of proliferating cells showing PCNA positivity and BrdU incorporation in the nuclei was decreased in a dose dependent manner by treatments of Paclitaxel and Cisplatin. Expression of type IV collagen also decreased after treatment with these two agents. Results of the TUNEL assay showed no change in the apoptosis of cells with regard to an increase in concentration of Paclitaxel and Cisplatin.
References
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Figure 1.
Morphological profiles of the transdifferentiated cells of the Paclitaxel-treated group (experimental group). Magnification: 200x. The group treated with 1 nM (B), 10 nM (C) and 100 nM Paclitaxel (D) showed slightly decreased cell number than control group (A).
Figure 2.
Morphological profiles of the transdifferentiated cells in the Cisplatin-treated group (experimental group II). Magnification: 200×. The group treated with 1 µM (B), 10 µM (C) and 100 µM Cisplatin (D) showed slightly decreased cell number than control group (A).
Figure 3.
The cell number of fibroblast per microscopic field during culture in experimental group I. The cell number was decreased by treatment of Paclitaxel in dose- * Significant decrease of the cell number dependent manner. in 100 nM Paclitaxel-treated group. (p<0.05)
Figure 4.
The cell number of fibroblast per microscopic field during culture in experimental group II. The cell number was decreased by treatment of Cisplatin in * Significant decrease of the cell dose-dependent manner. number in 100 µM Cisplatin-treated group. (p<0.05)
Figure 5.
Immunocytochemical detection of the incorporated BrdU in the nuclei of proliferating cells at day 7 in experimental group I. Magnification: 200×. The number of cells incorporated with BrdU in their nuclei decreased according to increasing concentrations of 1 nM (B), 10 nM (C) and 100 nM Paclitaxel (D) than control group (A).
Figure 6.
Immunocytochemical detection of the incorporated BrdU in the nuclei of proliferating cells at day 7 in experimental group II. Magnification: 200×. The number of cells incorporated with BrdU in their nuclei decreased according to increasing concentrations of 1 µM (B), 10 µM (C) and 100 µM Cisplatin (D) than control group (A).
Figure 7.
Immunocytochemical detection of the PCNA in the nuclei of proliferating cells at day 7 in experimental group I. Magnification: 200× The number of PCNA positive cells decreased according to increasing concentrations of 1 nM (B), 10 nM (C) and 100 nM Paclitaxel (D) than control group (A).
Figure 8.
Immunocytochemical detection of the PCNA positive proliferating cells on the cross section of cultured capsular bag at day 7 in experimental group I. PCNA positive cell (red arrow, a reddish brown diffuse or granular nucleus staining) was noted in control (A) (×200). PCNA positive cell was not noted in 1 nM (B), 10 nM (C) and 100 nM Paclitaxel (D) (×100).
Figure 9.
Immunocytochemical detection of the PCNA in the nuclei of proliferating cells at day 7 in experimental group II. Magnification:200× The number of PCNA positive cells(a reddish brown diffuse or granular nucleus staining) decreased according to increasing concentrations of 1 µM (B), 10 µM (C) and 100 µM Cisplatin (D) than control group (A).
Figure 10.
Immunocytochemical detection of the PCNA positive proliferating cells on the cross section of cultured capsular bag at day 7 in experimental group II. PCNA positive cell (red arrow) was noted in control (A) (×200). PCNA positive cell was not noted in 1 µM (B), 10 µM (C) and 100 µM Cisplatin (D) (×100).
Figure 11.
TUNEL staining analysis in experimental group I. Magnification:400× Apoptotic body (red arrow) was seen in control group (A). The number of apoptotic bodies showed no significant change according to increasing concentrations of Paclitaxel (1 nM (B), 10 nM (C), 100 nM (D)).
Figure 12.
TUNEL staining analysis in experimental group II. Magnification: 400× Apoptotic body (red arrow) was seen in control group (A). The number of apoptotic bodies showed no significant change according to increasing concentrations of Cisplatin (1 µM (B), 10 µM (C), 100 µM (D)).
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