Different temperature hyperthermia on human cervical cancer cell apoptosis and proliferation

Title: Different temperature hyperthermia on human cervical cancer cell apoptosis and proliferation Author: Zhou Jumei Degree-granting units: Central South University Keywords: hyperthermia;; Caski;; apoptosis;; cell proliferation;; cell cycle;; PCNA protein; hyperthermia;; Caski fine
Cell;; Smac / DIABLO;; survivin;; Neodymium Magnets caspase-3;; HPV16E6;; p53; magnetic fluid;; hyperthermia;; cervical cancer;; animal models;;
Cell proliferation Abstract: Background Cervical cancer is the female reproductive system, the highest incidence of cancer, and high-risk HPV infection is directly related to recent
To the incidence is rising and dropping. Advanced cervical cancer with radiotherapy and chemotherapy based. Because chemotherapy resistance and toxicity of chemotherapy and anti-
Should, advanced cervical cancer 5-year survival rate was 50%. Hyperthermia caused apoptosis and necrosis, both radiotherapy and chemotherapy sensitizing advantage in the Palace
Comprehensive treatment of cervical cancer increased 5-year survival and local control rates, while long-term toxicity was not increased. Hyperthermia on cervical cancer
Mechanisms remain unclear. 43 ℃ traditional hyperthermia that is most critical point of apoptosis, recent studies have found, 43 ℃ hot
Therapy is not for all types of cancer all have in vitro. The sensitivity of cells to hyperthermia and cell types and genetic characteristics of the
. Explore different cell lines contribute to the sensitivity of clinical hyperthermia hyperthermia individualized program. Hyperthermia is an emerging magnetic hyperthermia
Means, as compared with http://www.everbeenmagnet.com/en/products/110-sintered-neodymium-magnets conventional hyperthermia has the advantage of targeting positioning of heating, while the surrounding normal tissue damage, in the forefront of foreign
Adenocarcinoma Ⅱ clinical trials showed good efficacy, but have not been reported for cervical cancer. Therefore, we use
Heated at different temperatures cervical cancer cells in vivo to explore the different temperature hyperthermia on apoptosis and proliferation, as well as the intrinsic role of apoptosis
Mechanism for the clinical application of hyperthermia in cervical cancer provide some theoretical basis. Chapter temperature hyperthermia in cervical cancer
Caski apoptosis and proliferation Objective To investigate the temperature Caski hyperthermia in cervical cancer cell apoptosis, proliferation, and fine
Cell cycle. Methods Caski water bath for cervical cancer cell line 43 ℃, 45 ℃, 47 ℃ and 37 ℃ experimental group the control group hyperthermia,
Tetrazolium blue (MTT) to detect cell proliferation rate, apoptosis rate by flow cytometry, necrosis and cell cycle changes
, Immunohistochemical detection of proliferating cell nuclear antigen (PCNA) expression. Results of 45 ℃, 47 ℃ group of cell proliferation rate in 24h,
48h, 72h, 96h were 76.11 ± 5.32%, 81.08 ± 4.03%, 74.71 ± 3.78%, 72.16 ± 4.75% and 88.57 ±
3.77%, 91.33 ± 2.43%, 91.09 ± 1.17%, 92.05 ± 1.67%, significantly higher than 43 ℃ group, 26.57 ± 3.46%, 25.49 ±
2.76%, 22.80 ± 3.16%, 26.87 ± 4.01% (P <0.05). 45 ℃ and 47 ℃ group, cell proliferation was no significant difference between the rate of
Significant (P> 0.05). 45 ℃ group the highest rate of apoptosis, is 12.82 ± 1.77%, and other differences between the groups was significant
(P <0.05). 47 ℃ group cell necrosis highest rate of 39.15 ± 5.67%, the difference between the groups were significant (P <0.05). 43 ℃
, 45 ℃, 47 ℃ group G_1 phase cells than the control group decreased significantly (P <0.05), while the S-phase cells were significantly increased (P <0.05). Table of PCNA
Up with the hyperthermia temperature increases. 45 ℃, 47 ℃, the mean optical density (AOD) were 0.228 ± 0.022,0.214 ±
0.018, compared with the control group and 43 ℃ significant difference compared (P <0.05). Conclusion 45 ℃, and hyperthermia can significantly inhibit the above
System Caski cell proliferation, increased apoptosis and necrosis, cell cycle arrest in S phase, reducing the expression of PCNA. Chapter II
Hyperthermia-induced Smac / DIABLO expression of apoptosis promoting Caski Objective To study the cancer cells through different temperature heat Caski
After treatment of apoptosis-related genes p53, Smac / DIABLO, survivin, caspase-3 expression changes. Method using a water bath for
Caski cervical cancer cell line 43 ℃, 45 ℃, 47 ℃ hyperthermia 40min, real-time quantitative PCR after hyperthermia at different temperatures to continue to foster 2h
, 6h, 12h, 24h HPV16E6, p53, Smac / DIABLO, survivin, caspase-3 expression at the mRNA level.
Western Blotting detection of hyperthermia at different temperatures after 2h, 6h, 12h, 24h Smac / DIABLO, survivin, caspase-3 in
Changes in protein expression. Immunohistochemical detection of hyperthermia at different temperatures after 24h caspase-3 protein expression changes in cells.
Results (1) 43 ℃ at different time points after hyperthermia HPV16E6, Smac / DIABLO, survivin, caspase-3 mRNA expression than the control
Group increased (P <0.05). Compared with the control group, 43 ℃ 2h after hyperthermia decreased expression of survivin protein appeared, followed by survivin protein
Expression but increased (P <0.05). 43 ℃, and hyperthermia after 6h 24h Smac / DIABLO protein expression was significantly increased (P <0.05); and
caspase-3 protein expression increased after hyperthermia in 12-24h (P <0.05). (2) 45 ℃ at different time points after hyperthermia HPV16E6 mRNA,
survivin mRNA and protein expression than the control group decreased (P <0.05). 45 ℃ hyperthermia after 12-24h Smac / DIABLO and caspase-3
mRNA expression was significantly increased compared with the control group (P <0.05). 45 ℃ hyperthermia 6h Smac / DIABLO and caspase-3 protein expression on
Tune, and maintain to hyperthermia after 24h (P <0.05). (3) 47 ℃ at different time points after hyperthermia HPV16E6, Smac / DIABLO, survivin
mRNA expression was significantly lower than the control group (P <0.05); and caspase-3 mRNA in the 24h after hyperthermia compared with the control group increased (P <0.05).
47 ℃ hyperthermia 2h Smac / DIABLO protein expression and 2-6hcaspase-3 protein expression increased significantly compared with the control group (P <0.05), the
The latter two were significantly lower than the control group (P <0.05). 47 ℃ at all time points after hyperthermia survivin protein expression was significantly reduced or even
Loss (P <0.05). (4) 43 ℃, 45 ℃ and 47 ℃ hyperthermia wild-type p53 mRNA expression was significantly reduced compared with the control group (P <0.05).
(5) 43 ℃ and 45 ℃ hyperthermia 24h caspase-3 of the AOD values ​​than 37 ℃ and 47 ℃ was significantly increased (P <0.05). And 45 ℃ and 47 ℃ hyperthermia
Caspase-3 after the nuclear-positive index increased, and 37 ℃ and 43 ℃, the difference was significant (P <0.05). Conclusion Cervical cancer
Caski cells, Smac / DIABLO, survivn and caspase-3 mRNA and protein expression levels of temperature and cultured with hyperthermia
Between the. More than 45 ℃ hyperthermia inhibited HPV16E6 mRNA expression. Hyperthermia Smac / DIABLO Caski induced apoptosis pathway
May not rely on wild-type p53 gene regulation. Hyperthermia after caspase-3 translocation from the cytoplasm to the nucleus is an important matter of apoptosis
Pieces. Chapter temperature hyperthermia in human cancer xenografts of nude mice Objective To investigate the temperature hyperthermia
Human cervical carcinoma transplanted into nude mice role. Method employing cervical cancer Hela cell line established in nude mice xenograft model
Will Fe_3O_4 magnetic nanoparticles directly into the tumor tissue, in the alternating magnetic field for magnetic induction heating treatment temperature at 43 ℃, respectively,
And 47 ℃ last 30min, 3 days after repeated heating time, more 43 ℃ group hyperthermia group (43 ℃ MFH), 47 ℃ hyperthermia group (47 ℃ CMFH)
, A simple set of magnetic fluid (MF group), normal saline (NS group) and the control group the size of the tumor volume, tumor volume inhibition rate, survival and
Of PCNA-positive index. The results of the first 1-4 weeks after hyperthermia 47 ℃ MFH group tumor volume inhibition rates were 47.05%,
89.21%, 88.55%, 87.19%; 43 ℃ MFH group was 32.26%, 52.64%, 48.84%, 38.42%. Compared with the control group
Than, 43 ℃ MFH group and 47 ℃ MFH group can inhibit tumor growth (P <0.05). And 43 ℃ MFH group, 47 ℃ MFH group is more inhibitory effect
Significantly, there are 33.33% of the tumor regression rate (P <0.05). 43 ℃ MFH group and 47 ℃ MFH group, survival time was 36.67 ± 1.28d,
49.33 ± 3.19d, compared with the control group, 27.33 ± 1.41d significantly longer (P <0.05). And 47 ℃ MFH group and 43 ℃ MFH group's survival
The difference between also significant (P <0.05). Positive index of PCNA in 47 ℃ MFH group was significantly reduced to 12.33 ± 3.40%, and other groups
Compared, the difference was significant (P <0.05). Conclusion 43 ℃ MFH group and 47 ℃ MFH group can inhibit cancer transplanted into nude mice
Growth and prolong survival of nude mice, inhibited the proliferation of PCNA, but at 47 ℃ MFH group of more significant role, even up to cancer
Dissipated effect. Degree Year: 2009