| Research background:In cervical cancer, endometrial cancer and ovarian cancer, and said the three gynecological malignancies, the incidence of cervical cancer among the first, and most patients in developing countries, every year about 27 million people worldwide died of the disease, which gives women’s health and life posed a serious threat.Although cervical adenocarcinoma is rare in cervical cancer, but compared with squamous cell carcinoma, adenocarcinoma have a poor prognosis and high mortality,and the incidence of cervical adenocarcinoma in recent years compared with the previous increase and at a younger age trend.Because cervical adenocarcinoma radiosemsitivity low, so chemotherapy is attention.Currently cervical adenocarcinoma chemotherapy is still a platinum-based anticancer drugs, cisplatin-based past.And recent studies have found,compared with cisplatin,carboplatin was no significant difference in terms of the anti-tumor effect of cervical cancer, but its nephrotoxicity,gastrointestinal toxicity, neurotoxicity and other adverse reactions was significantly lower than cisplatin, so superior in cervical cancer chemotherapy.Carboplatin(CBP)belong to the second generation of platinum compound, was discovered in 1980, was approved for production and clinical application in 1990 in our country.Carboplatin is by changing the chemical structure of DNA of tumor cells and affect cell replication,which play anti-tumor effect.Although adverse reactions less than cisplatin, its side effects and drug resistance are still limits the clinical dose.Therefore, exploring new drugs for anti-tumor therapy or enhanced sensitivity to existing chemotherapy drugs has become a hot research in cervical adenocarcinoma chemotherapy.Rapamycin(RAPA) is a macrolide antibiotic.Initially rapamycin as a low toxicity antifungal studied, later it found its immune suppression and prevention of coronary restenosis after stent implantation and so on.While rapamycin for advanced kidney cancer, breast cancer, recurrent glioblastoma, osteosarcoma, advanced non-small cell lung cancer and other malignancies also have significant anti-tumor effect, and can increase the sensitivity of other anti-cancer drugs.Therefore, we hypothesized that rapamycin as an anti-cancer drugs for cervical adenocarcinoma also has killing effect,in its combination with carboplatin, the two have mutually reinforcing effect,it can increase the antitumor effect of carboplatin.Objective:View of rapamycin(RAPA) and carboplatin(CBP) under the drug alone and in combination on Hela cell proliferation, apoptosis, cell cycle arrest and other aspects of the impact, thus to provide a theoretical basis for seeking new drugs to cervical adenocarcinoma chemotherapy and increaseing existing chemotherapy drugs,chemosensitivity.Methods:(1) MTT staining detect the effect of rapamycin,carboplatin alone or in combination on the proliferation of cervical adenocarcinoma Hela cell.(2) Flow cytometry(FCM)PI staining detect the effect of rapamycin,carboplatin alone or in combination on the cell cycle distribution of cervical adenocarcinoma Hela cell.(3) Annexin V-7 AAD double staining detect the effect of rapamycin,carboplatin alone or in combination on the apoptosis of cervical adenocarcinoma Hela cell.(4) Reverse transcription- polymerase chain reaction(RT-PCR) technique detect the effect of rapamycin,、carboplatin alone or in combination on the Bcl-2 / Bax gene expression of cervical adenocarcinoma Hela cellResults:(1) MTT staining test results show:rapamycin or carboplatin alone can inhibit the proliferation of cervical adenocarcinoma Hela cells in a time- dependent manner,and the inhibitory effect of carboplatin is superior than rapamycin;the inhibitory effect of rapamycin in combination with carboplatin group is higher than rapamycin, carboplatin alone group, the differences were statistically significant(P<0.05).(2) Flow cytometry(FCM) PI staining test results show:Rapamycin or carboplatin alone can increase cervical adenocarcinoma Hela cells in G1 phase,reduce the proportion of cells in S phase and G2 phase in a time- dependent manner,and the effect of carboplatin is superior than rapamycin; the effect of rapamycin in combination with carboplatin group is superior than rapamycin,carboplatin alone group, the differences were statistically significant(P <0.05).(3) Annexin V-7 AAD double staining test results show:rapamycin or carboplatin alone can promote the apoptosis of cervical adenocarcinoma Hela cells in a time-dependent manner,and promoting effect of carboplatin is superior than rapamycin;the apoptosis rate of rapamycin in combination with carboplatin group is higher than rapamycin, carboplatin alone group, the differences were statistically significant(P<0.05).(4) Reverse transcription- polymerase chain reaction(RT-PCR) technology test results show:Rapamycin or carboplatin alone can make cervical adenocarcinoma Hela cells increased the expression of pro-apoptotic Bax gene, reduced the expression of anti-apoptotic Bcl-2 gene in a time- dependent manner,and the effect of carboplatin is superior to rapamycin; the effect of rapamycin in combination with carboplatin group is superior than rapamycin, carboplatin alone group, the differences were statistically significant(P <0.05).Conclusion:(1) Rapamycin or carboplatin alone can inhibite the proliferation of cervical adenocarcinoma Hela cells and induce the apoptosis of cervical adenocarcinoma Hela cells in a time- dependent manner, suggesting that both may have anti-tumor effect on cervical adenocarcinoma.(2) The effect of rapamycin in combination with carboplatin is superior than rapamycin, carboplatin alone,both in the inhibition of Hela cells proliferation and the promotion of Hela cells apoptosis induce a synergistic effect,suggesting that rapamycin can enhance the effect of chemotherapy carboplatin in cervical adenocarcinoma.(3) The antitumor mechanism and synergistic effect of rapamycin, carboplatin treating cervical adenocarcinoma Hela cells may related with blocking cell cycle progression, decreasing the expression of apoptotic genes Bcl-2/Bax. |
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