Appropriate empirical antifungal therapy is associated with a reduced mortality rate in intensive care unit patients with invasive fungal infection: A real-world retrospective study based on the MIMIC-IV database.

Objective: The study aimed to determine the prevalence and pathogens of invasive fungal infection (IFI) among intensive care unit (ICU) patients. The next goal was to investigate the association between empirical antifungal treatment and mortality in ICU patients. Methods: Using microbiological events, we identified all ICU patients with IFI and then retrieved electronic clinical data from the Medical Information Mart for Intensive Care IV (MIMIC-IV) database. The data were statistically analyzed using t-tests, chi-square tests, log-rank tests, and Cox regression. Results: The most commonly reported fungi were Candida (72.64%) and Aspergillus (19.08%). The most frequently prescribed antifungal medication was fluconazole (37.57%), followed by micafungin (26.47%). In the survival study of ICU patients and patients with sepsis, survivors were more likely to receive empirical antifungal treatment. In contrast, non-empirical antifungal therapy was significantly associated with poor survival in patients with positive blood cultures. We found that the current predictive score makes an accurate prediction of patients with fungal infections challenging. Conclusions: Our study demonstrated that empirical antifungal treatment is associated with decreased mortality in ICU patients. To avoid treatment delays, novel diagnostic techniques should be implemented in the clinic. Until such tests are available, appropriate empirical antifungal therapy could be administered based on a model that predicts the optimal time to initiate antifungal therapy. Additional studies should be conducted to establish more accurate predictive models in the future.

Individualized model for predicting COVID-19 deterioration in patients with cancer: A multicenter retrospective study.

The 2019 novel coronavirus has spread rapidly around the world. Cancer patients seem to be more susceptible to infection and disease deterioration, but the factors affecting the deterioration remain unclear. We aimed to develop an individualized model for prediction of coronavirus disease (COVID-19) deterioration in cancer patients. The clinical data of 276 cancer patients diagnosed with COVID-19 in 33 designated hospitals of Hubei, China from December 21, 2019 to March 18, 2020, were collected and randomly divided into a training and a validation cohort by a ratio of 2:1. Cox stepwise regression analysis was carried out to select prognostic factors. The prediction model was developed in the training cohort. The predictive accuracy of the model was quantified by C-index and time-dependent area under the receiver operating characteristic curve (t-AUC). Internal validation was assessed by the validation cohort. Risk stratification based on the model was carried out. Decision curve analysis (DCA) were used to evaluate the clinical usefulness of the model. We found age, cancer type, computed tomography baseline image features (ground glass opacity and consolidation), laboratory findings (lymphocyte count, serum levels of C-reactive protein, aspartate aminotransferase, direct bilirubin, urea, and d-dimer) were significantly associated with symptomatic deterioration. The C-index of the model was 0.755 in the training cohort and 0.779 in the validation cohort. The t-AUC values were above 0.7 within 8 weeks both in the training and validation cohorts. Patients were divided into two risk groups based on the nomogram: low-risk (total points ≤ 9.98) and high-risk (total points > 9.98) group. The Kaplan-Meier deterioration-free survival of COVID-19 curves presented significant discrimination between the two risk groups in both training and validation cohorts. The model indicated good clinical applicability by DCA curves. This study presents an individualized nomogram model to individually predict the possibility of symptomatic deterioration of COVID-19 in patients with cancer.

Thrombocytopenia in 737 adult intensive care unit patients: A real-world study of associated factors, drugs, platelet transfusion, and clinical outcome.

OBJECTIVE: We aimed to identify and represent factors associated with thrombocytopenia in intensive care unit, especially the pathogens and drugs related to severe and extremely thrombocytopenia. Then, we aim to compare the mortality of platelet transfusion and non-transfusion in patients with different degrees of thrombocytopenia. METHODS: We identified all thrombocytopenic patients in intensive care unit by using platelet-specific values and then extracted electronic health records from our Hospital Information System. Data were statistically analyzed with t test, chi-square test, and logistic regression. RESULTS: We found that infections (32.7%) were the most frequent cause associated with thrombocytopenia, followed by sepsis shock (3.93%) and blood loss (2.99%). Meanwhile, antifungals (p = 0.002) and bacterial infection (p = 0.037) were associated with severe and extremely severe thrombocytopenia. Finally, we found that the mortality of platelet transfusion and non-transfusion in patients was statistically significant for patients with platelet counts between 30 and 49/nL (χ2 = 9.719, p = 0.002). CONCLUSION: Infection and sepsis emerged as two primary factors associated with thrombocytopenia in intensive care unit. Meanwhile, antifungals and bacterial infection were associated with platelet counts less than 49/nL. Finally, platelet transfusion may be associated with reduced mortality in patients with platelet counts between 30 and 49/nL.

Advantages with prophylactic PEG-rhG-CSF versus rhG-CSF in breast cancer patients receiving multiple cycles of myelosuppressive chemotherapy: an open-label, randomized, multicenter phase III study.

BACKGROUND: PEG-rhG-CSF reduces neutropenia and improves chemotherapy safety. In China's registration trial (CFDA: 2006L01305), we assessed its efficacy and safety against rhG-CSF, and prospectively explored its value over multiple cycles of chemotherapy. METHODS: In this open-label, randomized, multicenter phase 3 study, breast cancer patients (n = 569) were randomized to receive PEG-rhG-CSF 100 µg/kg, PEG-rhG-CSF 6 mg, or rhG-CSF 5 µg/kg/d after chemotherapy. The primary endpoints were the incidence and duration of grade 3/4 neutropenia during cycle 1. Secondary endpoints included the incidence and duration of grade 3/4 neutropenia during cycles 2-4, the incidence of febrile neutropenia, and the safety. RESULTS: A once-per-cycle PEG-rhG-CSF at either 100 µg/kg or 6 mg was not different from daily injections of rhG-CSF for either incidence or duration of grade 3/4 neutropenia. Interestingly, a substantial difference was noted during cycle 2, and the difference became bigger over cycles 3-4, reaching a statistical significance at cycle 4 in either incidence (P = 0.0309) or duration (P = 0.0289) favoring PEG-rhG-CSF. A significant trend toward a lower incidence of all-grade adverse events was noted at 129 (68.98%), 142 (75.53%), and 160 (82.47%) in the PEG-rhG-CSF 100 µg/kg and 6 mg and rhG-CSF groups, respectively (P = 0.0085). The corresponding incidence of grade 3/4 drug-related adverse events was 2/187 (1.07%), 1/188 (0.53%), and 8/194 (4.12%), respectively (P = 0.0477). Additionally, PFS in metastatic patients preferred PEG-rhG-CSF to rhG-CSF despite no significance observed by Kaplan-Meier analysis (n = 49, P = 0.153). CONCLUSIONS: PEG-rhG-CSF is a more convenient and safe formulation and a more effective prophylactic measure in breast cancer patients receiving multiple cycles of chemotherapy.

Computational analysis reveals microRNA-mRNA regulatory network in esophageal squamous cell carcinoma.

MicroRNAs (miRNAs) are known to regulate post-transcriptional gene expression. They are involved in carcinogenesis and tumor progression. The aim of this study was to explore the microRNA-mRNA regulatory network in esophageal squamous cell carcinoma (ESCC) using comprehensive computational approaches. In this study we have selected a total of 11 miRNAs from one previously reported study in ESCC. The mRNA targets of these miRNAs were predicted using various algorithms. The expression profiles of these mRNA targets were identified on DNA microarray experiment dataset across ESCC tissue samples. Based on the miRNA-mRNA regulatory relationships, the network was inferred. A total of 23 miRNA-mRNA regulatory interactions, with 11 miRNAs and 13 mRNA targets, were inferred in ESCC. The miRNA-mRNA regulatory network with increased confidence provides insights into the progression of ESCC and may serve as a biomarker for prognosis or the aggressiveness of ESCC. However, the results should be examined with further experimental validation.

[Pollutional Characteristics and Sources Analysis of Polycyclic Aromatic Hydrocarbons in Atmospheric Fine Particulate Matter in Lanzhou City].

Polycyclic aromatic hydrocarbons (PAHs) are a group of important toxic compounds. In order to detect the pollutional characteristics of atmospheric PAHs in Fine Particulate Matter (PM2.5), a total of 60 PM2.5 samples were collected in Lanzhou City during the winter of 2012 and summer of 2013. The GC/MS measurement results of the samples demonstrated the averagely total mass concentrations of the most significant 16 homologues of PAHs were (191.79±88.29) ng·m-3 and (8.94±4.34) ng·m-3 in winter and summer respectively, indicating a higher pollution level in winter. In winter, the snowfall was the most important meteorological factor for the decrease of PAHs mass concentration in PM2.5. The percentages of PAHs with 4 rings were the highest in both winter (51.40%) and summer (49.94%) in Lanzhou. The percentage of PAHs with 5-6 rings in summer (41.04%) was higher than that in winter (24.94%). However, the percentage of PAHs with 2-3 rings in summer (9.03%) was lower than that in winter (23.67%). Based on the analysis of characteristic ratios, we concluded that the PAHs in atmospheric PM2.5 in Lanzhou were mainly sourced from coal and vehicle emissions in winter, especially the diesel vehicles. The absolute contributions of all possible PAHs pollution sources were insignificant in summer, with relatively higher contribution from gasoline vehicles.

New dating of the Homo erectus cranium from Lantian (Gongwangling), China.

The Homo erectus cranium from Gongwangling, Lantian County, Shaanxi Province is the oldest fossil hominin specimen from North China. It was found in 1964 in a layer below the Jaramillo subchron and was attributed to loess (L) L15 in the Chinese loess-palaeosol sequence, with an estimated age of ca. 1.15 Ma (millions of years ago). Here, we demonstrate that there is a stratigraphical hiatus in the Gongwangling section immediately below loess 15, and the cranium in fact lies in palaeosol (S) S22 or S23, the age of which is ca. 1.54-1.65 Ma. Closely spaced palaeomagnetic sampling at two sections at Gongwangling and one at Jiacun, 10 km to the north, indicate that the fossil layer at Gongwangling and a similar fossil horizon at Jiacun were deposited shortly before a short period of normal polarity above the Olduvai subchron. This is attributed to the Gilsa Event that has been dated elsewhere to ca. 1.62 Ma. Our investigations thus demonstrate that the Gongwangling cranium is slightly older than ca. 1.62 Ma, probably ca. 1.63 Ma, and significantly older than previously supposed. This re-dating now makes Gongwangling the second oldest site outside Africa (after Dmanisi) with cranial remains, and causes substantial re-adjustment in the early fossil hominin record in Eurasia.

Intravenous flurbiprofen axetil enhances analgesic effect of opioids in patients with refractory cancer pain by increasing plasma ß-endorphin.

BACKGROUND: The study aimed to investigate the analgesic effect of a combination of intravenous flurbiprofen axetil and opioids, and evaluate the relationship between refractory pain relief and plasma ß-endorphin levels in cancer patients. MATERIALS AND METHODS: A total of 120 cancer patients was randomly divided into two groups, 60 patients took orally morphine sulfate sustained-release tablets in group A, and another 60 patients receiving the combination treatment of intravenous flurbiprofen axetil and opioid drugs in group B. After 7 days, pain relief, quality of life improvement and side effects were evaluated. Furthermore, plasma ß-endorphin levels were measured by radioimmunoassay. RESULTS: With the combination treatment of intravenous intravenous flurbiprofen axetil and opioids, the total effective rate of pain relief rose to 91.4%, as compared to 82.1% when morphine sulfate sustained-release tablet was used alone. Compared with that of group A, the analgesic effect increased in group B (p=0.031). Moreover, satisfactory pain relief was associated with a significant increase in plasma ß-endorphin levels. After the treatment, plasma ß-endorphin level in group B was 62.4±13.5 pg/ml, which was higher than that in group A (45.8±11.2 pg/ml) (p<0.05). CONCLUSIONS: Our results suggest the combination of intravenous flurbiprofen axetil and opioids can enhance the analgesic effect of opioid drugs by increasing plasma ß-endorphin levels, which would offer a selected and reliable strategy for refractory cancer pain treatment.

Co-transduction of ribosomal protein L23 enhances the therapeutic efficacy of adenoviral-mediated p53 gene transfer in human gastric cancer.

Induction of murine double minute 2 (MDM2) expression is thought to be a determinant of resistance to p53 gene therapy for cancer. Previous studies have revealed that ribosomal protein L23 (RPL23) inhibits MDM2-mediated p53 degradation through direct binding to MDM2. In addition, ectopically expressed RPL23 was reported to interact with MDM2 in both the nucleus and cytoplasm, by which RPL23 indirectly inhibited MDM2-p53 binding. Based on the known molecular properties of the RPL23 protein, it was speculated that co-transduction of RPL23 may protect wild­type p53 protein from MDM2-mediated inactivation and, thus, improve the effect of delivering therapeutic exogenous p53. To test this hypothesis, we constructed a bicistronic adenoviral vector expressing both the RPL23 and p53 genes (Ad-RPL23/p53) and compared its tumor-suppressor activity in human gastric cancer with that of a single gene vector for p53 (Ad-p53). In the in vivo and in vitro experiments, we observed that treatment with Ad-RPL23/p53 resulted in a stronger antitumor response compared to that obtained using Ad-p53. Moreover, the antitumor response of the bicistronic adenovirus was obtained not only in MGC803 cells (endogenous mutant p53) but also in MKN45 cells (endogenous wild­type p53) which were initially resistant to p53 gene transfer, indicating that co-transduction of RPL23 also expanded the utility of p53 gene therapy. Furthermore, in an orthotopic nude mouse model of human gastric cancer, we found that the survival benefit was greater after Ad-RPL23/p53 treatment than after Ad-p53. Taken together, the data presented here demonstrate that co-transduction of RPL23 enhances the therapeutic efficacy of adenoviral-mediated p53 gene transfer in models of human gastric cancer and support the use of this strategy for cancer treatment.

MiR-26a regulates cell cycle and anoikis of human esophageal adenocarcinoma cells through Rb1-E2F1 signaling pathway.

Resistance to anoikis, the subtype of apoptosis induced by lack of matrix adhesion, contributes to malignant transformation and development of metastasis. MicroRNAs play key regulatory roles in tumorigenesis and metastasis. In this study, we described that miR-26a, which is usually downregulated in tumor cells, is involved in the acquisition of anoikis-resistance of human esophageal adenocarcinoma (EA) cells. Results of qRT-PCR in clinical samples showed that downregulated miR-26a expression is related to tumorigenesis and metastasis of EA. In vitro experiments determined that miR-26a directly participates in the regulation of cell cycle and anoikis of human EA OE33 cells. Further, we identified that Rb1 is the direct functional target of miR-26a, and revealed that the reduction of miR-26a expression leads to increased Rb1 protein level and thus inhibits the function of E2F1, by which it influences the phenotypes of cell cycle and anoikis. The findings we reported here presented the evidence that miR-26a may be involved in regulation of anoikis-resistance of EA cells. Targeting miR-26a may provide a novel strategy to inhibit metastasis.

Peritumoral lymphatic microvessel density is related to poor prognosis in lung adenocarcinoma: A retrospective study of 65 cases.

Although recent investigations have identified that lymphangiogenesis is associated with regional lymph node metastasis and tumor prognosis in non-small cell lung cancer (NSCLC), peritumoral lymphatic microvessel density (LMVD) and its prognostic significance in lung adenocarcinoma remain unknown. In the present study, we assessed peritumoral LMVD in lung adenocarcinoma and investigated its correlation with patient prognosis. Using immunohistochemistry (SP method), the D2-40-positive peritumoral LMVD count in lung adenocarcinoma was found to be 11.56±10.73, which was higher than intratumoral LMVD (P<0.001), and was found to be associated with lymphatic metastasis (P=0.003) and pTNM staging (P=0.046). Furthermore, a significant difference in the patient overall survival time was demonstrated between tumors with a high peritumoral LMVD and those with a low peritumoral LMVD (P=0.005). Finally, using multivariate analysis, it was determined that peritumoral LMVD, lymphatic metastasis and pTNM staging were independent prognostic factors. In conclusion, the results suggest that D2-40-positive peritumoral LMVD may predict the prognosis of lung adenocarcinoma.

Giant adenocarcinoma of unknown origin penetrating from the pelvic cavity to subcutaneous tissue: A case report and review of the literature.

In this study, we present a case of a 43-year-old female patient who was admitted to our hospital due to a giant mass on the left buttock. Imaging tests revealed that the mass was a solid-cystic tumor with a large size of 143×430×180 mm, penetrating from the pelvic cavity to the subcutaneous tissue. Pathology tests indicated a metastatic mucinous adenocarcinoma which was most likely of gastrointestinal origin. However, there was no evidence to confirm the existence of malignant changes in the gastrointestinal tract.

Tumor-associated macrophages infiltration is associated with peritumoral lymphangiogenesis and poor prognosis in lung adenocarcinoma.

Tumor-associated macrophages (TAMs) have been implicated in promoting tumor progression. Nowadays, adenocarcinoma has surpassed squamous cell carcinoma as the most frequent type of lung cancer, but in lung adenocarcinoma, the correlation of TAMs with lymphangiogenesis patients remains unclear. The aim of this study was to examine the relationship between TAMs and lymphangiogenesis and the lung adenocarcinoma patients' prognosis. Tumor specimens from 65 patients with lung adenocarcinoma were determined for TAMs count and lymphatic microvessel density (LMVD) by immunohistochemistry. A positive correlation existed between TAMs count and D2-40-positive peritumoral LMVD (r=0.069, P<0.001). TAMs infiltration was significantly associated with P-TNM staging (P=0.042) and lymph node metastasis (P=0.037), and peritumoral LMVD was correlated with lymph node metastasis (P=0.003). A significant difference in overall survival was detected not only between tumors with a high TAMs count and a low TAMs count (P=0.009) but also between tumors with a high peritumoral LMVD and a low peritumoral LMVD (P=0.005). Both TAMs count and peritumoral LMVD were independent prognostic factors for overall survival. Our results indicate that TAMs infiltration correlates with tumor lymphangiogenesis and poor survival in lung adenocarcinoma.

[Efficacy of NO regimen and NP regimen on advanced non-small cell lung cancer: a prospective randomized trial].

BACKGROUND & OBJECTIVE: Oxaliplatin (LOHP) is an effective drug in treatment of non-small cell lung cancer (NSCLC) with mild toxicities to gastrointestinal tract, kidney, and bone marrow. Cisplatin (DDP) plus vinorelbine (NVB) constitute the first-line regimen (NP regimen) for NSCLC. This study was to compare the short-term response, long-term outcome, and adverse events between advanced NSCLC patients received NO regimen (LOHP plus NVB) and NP regimen. METHODS: A total of 90 patients with advanced NSCLC were randomized into NO group (58 patients, 25 mg/m(2) of NVB, day 1 and day 8; 130 mg/m(2) of LOHP, day 1) and NP group (32 patients, 25 mg/m(2) of NVB, day 1 and day 8; 50 mg/m(2) of DDP, day 2 and day 3). The short-term response, long-term outcome, adverse events, and survival status of the 2 groups were observed. RESULTS: The response rates were 33.33% in NO group, and 35.48% in NP group, but no significant difference was detected between the 2 groups (P > 0.05). The clinical benefit response rate was significantly higher in NO group than in NP group (80.70% vs. 64.52%, P < 0.05). The median time to progression (TTP) was 17 weeks in NO group, and 15 weeks in NP group; the median time of remission was 21 weeks in NO group, and 19 weeks in NP group; the median survival time was 39 weeks in NO group, and 37 weeks in NP group; the 1-year survival rate was 37.93% in NO group, and 31.25% in NP group. No significant differences were detected between the 2 groups. The incidence rates of phlebitis and grade I-II peripheral neuritis were significantly higher in NO group than in NP group (77.59% vs. 50.00%, P<0.01; 43.10% vs. 15.63%, P<0.01). The incidence rate of grade III-IV nausea/vomiting was significantly higher in NP group than in NO group (31.25% vs. 3.45%, P<0.05). CONCLUSIONS: The efficacy of NO regimen on advanced NSCLC is similar to that of NP regimen, but the clinical benefit response rate is higher in NO group than in NP group. In short, NO regimen may be recommended as the first-line chemotherapy regimen for advanced NSCLC.

[Radiation sensitizing effect of anti HPV16E6-ribozyme on cervical carcinoma cell line].

BACKGROUND & OBJECTIVE: Human papillomavirus (HPV) are the most important etiologic factor for cervical carcinoma. E6 virus gene is one of the most important oncogene and expression levels of E6 transforming oncoproteins of high risk HPV genotypes, such as HPV16, appear to be necessary for maintaining the malignant phenotype. Radiation treatment represents a standardized and effective modality for contemporary cervical carcinoma therapy. The goal of this study was to investigate the radiation sensitizing effect of anti-HPV16 E6-ribozyme on cervical carcinoma cell line. METHODS: With the method of lipofectin transfection, the anti-HPV16 E6 ribozyme and empty eucaryotic expressing plasmids were transfected into CaSKi cell line, which named as CaSKi-R and CaSKi-P, respectively. The expression of ribozyme in transfected cells was observed by RNA dot blot. The amounts of E6 mRNA in the three kinds of cells were detected by Northern blot. The growth rates of the CaSKi and transfected cells were examined by cell count and their sensitivity to radiotherapy were examined by colony formation test. The apoptosis rates of each cell was determined by PI/Annexin V stained methods. Expressions of p53, bcl-2, and bax were determined by Flow cytometry analysis. RESULTS: Anti-HPV16 E6-ribozyme can be expressed stably in transfected CaSKi-R cells. Northern blot showed that E6 mRNA was less in CaSKi-R than in CaSKi and CaSKi-P. The growth rate of CaSKi-R was much slower than that of CaSKi and CaSKi-P. The sensitivity of CaSKi-R cells to radiotherapy increased more than that of CaSKi and CaSKi-P cells. The ability of colony formation decreased (P < 0.05), while the apoptosis rates of CaSKi-R cells increased more than that of CaSKi and CaSKi-P cells(P < 0.01). Anti-HPV16 E6-ribozyme did significantly upregulate expression of p53, bax protein, and downregulate the expression of bcl-2 protein before and after radiotherapy (P < 0.01). CONCLUSION: CaSKi-R cells transfected with Anti-HPVE 6-rivozyme showed growth inhibition and increased sensitivity to radiotherapy.

[Effects of lipofectin-mediated alpha1,4Gal T antisense oligonucleotide on human glioma cell line SWO-38].

OBJECTIVE: To study the effects of alpha1,4Gal T antisense oligonucleotide mediated by lipofectin on human glioma cell line SWO-38. METHODS: SWO38 glioma cells were exposed to 10 micromol/L alpha1,4Gal T antisense oligonucleotide for 72 h by means of lipofectin transfection, and the growth inhibition of the cells was detected by colony-forming unit assay. Analysis of DNA fragmentation was performed with flow cytometry (FCM) and agarose gel eletrophoresis with Fas protein expression determined by FCM. RESULTS: alpha1,4Gal T antisense oligonucleotide significantly inhibited the growth of glioma cells (P<0.01) and induced apoptosis in SWO-38 cells (P<0.05). Marked up-regulation of Fas protein expression was observed in response to the treatment (P<0.01). CONCLUSION: alpha1,4Gal T antisense oligonucleotide can significantly inhibit SWO-38 cell proliferation and induce cellular apoptosis, the mechanism of which may involve up-regulated Fas expression.

[Effect of anti-HPV16 E6-ribozyme on the sensitivity of cervical carcinoma cell line to chemotherapy].

OBJECTIVE: To study the role of anti-HPV16E6-ribozyme in modulating the sensitivity of cervical carcinoma cell line to chemotherapy. METHODS: By way of lipofectin transfection, anti-HPV16E6-ribozyme antibody and empty eukaryotic expression plasmids were respectively transfected into CaSKi cells that were designated as CaSKi-R and CaSKi-P cells accordingly. Ribozyme expression in the transfected cells was subsequently observed with RNA dot blot, and the amount of E6 mRNA expression detected by Northern blotting. Cell count was performed for determining the growth rate of the non-transfected and transfected CaSKi cells and colony formation test was employed to examine the sensitivity of the cells to chemotherapy. PI/Annexin V staining was conducted to determine the apoptosis rates of each cell line in response to chemotherapy. RESULTS: As shown by dot blot analysis, stable expression of anti-HPV16E6-ribozyme was achieved in CaSKi-R cells, in which lower E6 mRNA expression level was observed by means of Northern blotting in comparison with those in CaSKi-P and CaSKi cells. Significant slow-down of the growth rate occurred in CaSKi-R cells as compared with the growth rate of the other 2 cell lines, but no differences in relative cloning efficiency and apoptosis rates of the 3 cell lines were observed in response to taxol treatment (P>0.05), while cis-platinum treatment produced comparatively deceased relative cloning efficiency and increased apoptosis rate in CaSKi-R (P<0.01). CONCLUSION: Transfection by anti-HPVE6-ribozyme may inhibit the growth of CaSKi cells and increase their sensitivity to cis-platinum but not to taxol.

Effects of anti-HPV16 E6-ribozyme on the proliferation and apoptosis of human cervical cancer cell line CaSKi.

OBJECTIVE: To study the characterization of anti-HPV16E6-ribozyme transfected into cultured human cervical cancer cell line, and to investigate the effect of ribozyme on the proliferation and apoptosis of the cells. METHODS: By way of lipofectin transfection, anti-HPV16E6-ribozyme, a ribozyme designed to specifically cleave the HPV16E6 gene, and empty vector expression plasmids were respectively transfected into CaSKi cells which were subsequently designated as CaSKi-R and CaSKi-P accordingly. The expression of ribozyme in the transfected cells was observed by RNA dot blot analysis, and E6 mRNA expression was detected by Northern blotting in the 3 kinds of cells whose growth curves and clone-forming ability on soft agar were studied, with their tumorgenicity observed by percutaneous inoculation of the cells in nude mice. Flow cytometry was employed to assess the apoptosis rates and the expression of the genes including c-myc, bcl-2, p53, and fas etc. RESULTS: Stable expression of anti-HPV16E6-ribozyme was observed in CaSKi-R cells that had less E6 mRNA expression than CaSKi had as shown by Northern blotting. When compared with those of CaSKi cells, the growth rate and clone-forming ability on soft agar of CaSKi-R were reduced, while those of CaSKi-P evinced no significant changes. The tumorgenicity of CaSKi-R in nude mice was also comparatively decreased, with markedly elevated apoptosis rate. Anti-HPV16E6-ribozyme reduced the expressions of E6, c-myc, bcl-2, PCNA and C-erbB2 genes but increased p53 expression in CaSKi-R cells, which, however, did not happen in CaSKi-P cells. The expression of Fas underwent no significant changes in response to the transfection. CONCLUSION: Anti-HPVE6-ribozyme inhibits proliferation and induces apoptosis in CaSKi cells, possibly due to the decrease of E6 gene expression that triggers a series of changes in the gene expressions of the cells.